INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL Do d
MANUAL TECNICO INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL TMs: Leonor Armanet, Juana Vargas, Amalia Cárcamo, Carolina Hernández
Ste
2012
CURSO 2015 Actualizado por TMs Josefina Josefin a Barrera y Catalina Salinas Sali nas
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
INTRODUCCION La Medicina Transfusional es un área de la Medicina que ha sufrido diversos y rápidos cambios a lo largo de su historia, en especial en los últimos años, lo que constituye un constante desafío para los que trabajan en ésta área. Hemos desarrollado este texto que pretende servir de guía tanto a estudiantes de la Carrera de Tecnología Médica como a profesionales que trabajan en Servicios de Sangre. En
el
presente
Manual
se
describen
aspectos
básicos
de
Inmunohematología y Medicina Transfusional, se describen las técnicas y procedimientos que se utilizan con mayor frecuencia en una Unidad de Medicina Transfusional y en un Centro de Sangre. Se incorpora un cuestionario de autoevaluación al término de cada una de las unidades temáticas, que permite que el estudiante pueda conocer el nivel de comprensión y manejo de las materias que aquí tratamos. Pretendemos además hacer reflexionar a quienes se inician en esta disciplina sobre aspectos relevantes del quehacer del Tecnólogo Médico en Medicina Transfusional, en tópicos como control de calidad, metodologías, registros, etc., con el propósito de lograr en ellos un espíritu crítico y autonomía en su quehacer profesional.
Pretendemos reforzar el aprendizaje y
comprensión del texto con talleres dirigidos y discusión de casos, buscando a través del trabajo cooperativo las respuestas a las interrogantes que aquí planteamos, y así también fortalecer aspectos que pueden no estar explícitamente tratados en éste manual.
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INDICE UNIDAD DONACIÓN DE SANGRE SELECCION DE UN DONANTE DE SANGRE
1.- INTRODUCCION: 2.- EL LLENADO DE D E LA FICHA DE DONANTE Y EL E L CUESTIONARIO: 3.- SELECCION DEL DONANTE: 4.- CONDICIONES DE LA ENTREVISTA ENTREV ISTA DEL DONANTE DONA NTE DE SANGRE SANGR E 5.- PRINCIPIOS DE UNA ENTREVISTA: ENTREVIS TA: 6.- CRITERIOS DE SELECCIÓN PREGUNTAS A REALIZAR DURANTE REALIZAR DURANTE LA ENTREVISTA AL DONANTE PROCEDIMIENTOS A UTILIZAR EN UTILIZAR EN LA ATENCION Y SELECCION DE UN DONANTE DE SANGRE I.- DETERMINACION DE Hb UTILIZANDO SULFATO DE COBRE. II.-DETERMINACION II.-DETERMINAC ION DE HEMOGLOBINA EN DONANTES CON EL METODO HEMOCUE III.- TECNICA EN LAMINA PARA CONTROLAR ANTIGENOS ABO EN GLOBULOS ROJOS. IV.- CONTROL DE CALIDAD DEL PROCEDIMIENTO DE EXTRACCION EN UN DONANTE 1.- CULTIVO BACTERIOLÓGICO DEL SITIO DE PUNCIÓN. 2.- CULTIVO BACTERIOLÓGICO DE LAS UNIDADES DE SANGRE. 3.- CONTROL DE CALIDAD DEL VOLUMEN DE SANGRE EXTRAÍDO A CADA DONANTE. V.- MEDICION DE LA PRESION ARTERIAL VI.- PREPARACION DE LA ZONA DE PUNCION VENOSA. VII.- PUNCION VENOSA Y EXTRACCION DE SANGRE. VIII.- ATENCION DEL DONANTE DESPUES DE LA EXTRACCION REACCIONES ADVERSAS A LA DONACION DE SANGRE I.- REACCIONES LOCALES: II.-REACCIONES GENERALIZADAS:
6 7 7 8 8 8 9 11 12 21 21 22 24 26 26 26 26 27 28 29 31 31 31 32
REGISTROS
34
ALMACENAMIENTO ALMACENAMIEN TO Y TRANSPORTE TRANSPOR TE DE HEMOCOMPONENTES HEMOCO MPONENTES GESTION DE STOCK EN STOCK EN BANCO DE SANGRE
37 37
UNIDAD INMUNOHEMATOLOGIA
INTRODUCCION A LA INMUNOHEMATOLOGIA REACTIVOS BASICOS A USAR EN USAR EN INMUNOHEMATOLOGIA Y BANCO DE SANGRE 1.- BUFFER FOSFATO Y BUFFER FOSFATO SALINO 2.- PREPARACION PREPARAC ION DE SUERO AB A B INERTE 3.- MEDIO DE BAJA FUERZA IÓNICA (LISS): 4.- EDTA - SALINO SA LINO 5.- SOLUCION DE ALSEVER A LSEVER MODIFICADA: MODIF ICADA: 6.- CONVERSION DE PLASMA A SUERO TRATAMIENTO DE MUESTRAS CON COAGULACION INCOMPLETA PREPARACION DE LECTINAS PARA USO EN INMUNOHEMATOLOGIA REACCION DE CELULAS POLIAGLUTINABLES CON LECTINAS LAVADO DE ERITROCITOS Y PREPARACION DE SUSPENSIONES CELULARES GRADUACION DE LOS RESULTADOS INMUNO-HEMATOLOGICOS CALIBRACION DE CENTRIFUGAS SEROLOGICAS PARA INMUNOHEMATOLOGIA CARACTERISTICAS DE LOS SUEROS CLASIFICADORES.
44 44 46 48 49 49 50 50 51 51 52 53 53 54 56 58
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
DETERMINACION DE ANTIGENOS SOLUBLES ABH Y LEWIS EN SALIVA FRECUENCIAS GENICAS DE SISTEMAS SANGUINEOS EN DIFERENTES POBLACIONES CARACTERISTICAS GENERALES DE SISTEMAS SANGUINEOS DE IMPORTANCIA CLINICA TEST ANTIGLOGULINA HUMANA TEST ANTIGLOBULINA HUMANA DIRECTO (TAD) TEST DE COOMBS DIRECTO TEST ANTIGLOBULINA HUMANA INDIRECTO (TAI) TEST DE COOMBS INDIRECTO PREPARACION DE GLOBULOS ROJOS CONTROL PARA TEST ANTIGLOBULINA HUMANA CLASIFICACION R H (D): TÉCNICA EN TUBO CLASIFICACION DE GRUPO ABO Y R H: TÉCNICA EN MICROPLACA DETERMINACION DE GENOTIPO RH Frecuencia de los haplotipos Rh en población chilena. ALGORITMO A UTILIZAR EN UTILIZAR EN LA CLASIFICACIÓN R H D DETECCION DE ANTICUERPOS IRREGULARES: TÉCNICA EN TUBO DETECCION DE ANTICUERPOS IRREGULARES EN PAPAINA DETECCIÓN DE ANTICUERPOS EN MEDIO LISS DETECCIÓN DE ANTICUERPOS IRREGULARES: TÉCNICA EN COLUMNA IDENTIFICACION DE ANTICUERPOS IRREGULARES COMPORTAMIENTO SEROLOGICO DE LOS PRINCIPALES ANTICUERPOS IRREGULARES CALCULO DE PROBABILIDAD: TECNICA DE IDENTIFICACION DE AC. EN BROMELINA TITULACION DE ANTICUERPOS PRUEBA CRUZADA O CROSMATCH ESTANDARIZACION DEL MEDIO DE BAJA FUERZA IONICA (LISS) PREPARACION Y ESTANDARIZACION DE SOLUCIONES USO DE DTT PARA DIFERENCIAR ANTICUERPOS DIFERENCIAR ANTICUERPOS IGM DE IGG TRATAMIENTO DE ERITROCITOS CON DTT ABSORCION DE ANTICUERPOS ELUCION DE ANTICUERPOS ESTUDIO R H EN CELULAS CON TAD POSITIVO AUTOADSORCION DE ANTICUERPOS CALIENTES (ZZAP) ADSORCION DIFERENCIAL EN CALIENTE USANDO ZZAP DETERMINACION DE ESPECIFICIDAD DE AUTO ANTICUERPOS FRIOS DEMOSTRACION DE AUTOAGLUTININAS FRIAS (CRIOAGLUTININAS) DE ALTO TITULO ESTUDIO DE CALIDAD DE UN SUERO ANTIGLOBULINA HUMANA I.-ESPECIFICIDAD: II.-EVALUACIÓN DE LA ACTIVIDAD ANTICOMPLEMENTO ANTICOMPLEME NTO III.- POTENCIA Y SENSIBILIDAD DEL SUERO AGH: CARACTERISTICAS GENERALES DE METODOS USADOS EN INMUNOHEMATOLOGIA ERITROCITOS CONGELADOS PARA FINES DE LABORATORIO ESTUDIO INMUNOHEMATOLÓGICO DE UNA MUESTRA
SANGRE Y COMPONENTES SANGUINEOS PREPARACION Y CONSERVACION DE COMPONENTES SANGUINEOS CONTENIDO DE LAS SOLUCIONES PRESERVADORAS PRESERVAD ORAS (g/L) CAMBIOS BIOQUIMICOS DE SANGRE CONSERVADA CONSERVAD A CON CPD Y CPDA-1 PREPARACION DE COMPONENTES SANGUINEOS POR CENTRIFUGACIÓN. POR CENTRIFUGACIÓN. CARACTERISTICAS CARACTERIS TICAS DE LOS PRINCIPALES COMPONENTES SANGUINEOS CONTENIDO APROXIMADO DE LEUCOCITOS EN LOS COMPONENTES SANGUINEOS REDUCCION DE LEUCOCITOS EN LOS COMPONENTES SANGUINEOS:
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A.- REACCIONES TRANSFUNSIONALES TR ANSFUNSIONALES INMEDIATAS B.- REACCIONES TRANSFUSIONALES RETARDADAS CUADRO RESUMEN DE LOS EFECTOS ADVERSOS DE LA TRANSFUSIÓN SANGUÍNEA INVESTIGACION DE UNA REACCION TRANSFUSIONAL
ENFERMEDADES ENFERMEDADES DE TRANSMISION SANGUINEA TESTS SEROLOGICOS EN EL DIAGNOSTICO DE HEPATITIS VIRAL CAUSAS POSIBLES DE RESULTADOS FALSOS EN TEST ELISA EVALUACION DE PROCEDIMIENTOS DE TAMIZAJE O SCREENING
BIBLIOGRAFIA
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141 142 142 143
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
UNIDAD DONACIÓN DE SANGRE
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
SELECCION DE UN DONANTE DE SANGRE 1.- INTRODUCCION: La seguridad de la transfusión de sangre o de sus componentes comienza con la apropiada selección del donante. Para lograr este objetivo se dispone de una Ficha de Donante y un Cuestionario para aplicar durante la entrevista a los posibles donantes, con la que pretendemos lograr una correcta selección de ellos, apoyándose además con folletos informativos (Ver anexos al término de la unidad) acerca de las enfermedades que se transmiten por la sangre y de la seguridad de la extracción sanguínea. Es necesario enfatizar también la necesidad de ofrecer al donante un proceso seguro y controlado. El proceso de atención de donantes de sangre consta de las siguientes etapas: a.- Recepción, identificación y codificación del donante: La recepción debe ser cálida. Solicite la información necesaria para identificar con seguridad al donante y carnet de identidad o documento con RUT y foto, llenando con el máximo de precisión todos los datos que aparecen en la Ficha de Donante de Sangre, e ingresándolos al sistema informático. El sistema asignará un número único de donación y emitirá etiquetas con código de barras, que deberán adherirse a los registros, bolsas y tubos. b.- Verificación del estado general del donante: Incluye peso, pulso, presión arterial, hemoglobina y aspecto general del donante, información que también debe registrar en la Ficha de Donante de Sangre. c.- Entrevista y aplicación del cuestionario:comprende una serie de preguntas específicas que tienen por objeto recoger una clara información para que determine si el donante cumple o no con los requisitos exigidos para donar. La duración de este procedimiento es de 8 a 10 minutos. d.- Compromiso del donante: Todo donante debe firmar el Consentimiento Informado contenido en la Ficha de Donante de Sangre, declarando entender las preguntas formuladas, que la información entregada es verdadera yque autoriza el procedimiento. e.- Agradecimiento por la donación e invitación a ser donante voluntario: Usted debe
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
2.- EL LLENADO DE LA FICHA DE DONANTE Y EL CUESTIONARIO: a.- Quien: El Tecnólogo Médico responsable de la sección. b.- Como: Preguntando al donante uno a uno los aspectos que contiene la "Ficha de Donante de Sangre" y el “Cuestionario”, recogiendo la información pertinente, teniendo presente la importancia de explicar en términos simples cada punto y de ganar la confianza del donante para lograr respuestas verdaderas. La entrevista que se realiza para llenar el cuestionario debe ser confidencial. c.-Cuando: Una vez que el posible donante ha leído la información acerca de las infecciones de transmisión sanguínea, tendientes a lograr la autoexclusión y se ha interiorizado sobre el procedimiento de la extracción (Revisar folletos relativos al tema).
3.- SELECCION DEL DONANTE: a.- Se entrevistará al donante de sangre, utilizando un cuestionario basado en las normas de selección establecidas en el país. b.- Se evaluará la información recogida, determinando la aceptación o exclusión del posible donante. c.- En caso de exclusión se explicará al donante en la forma más clara y completa posible, si la causa es definitiva o transitoria, se dará las recomendaciones pertinentes, verificando que el donante ha comprendido la situación, y se registrará la causa en la ficha del donante, utilizando el código respectivo. d.- Si no hay motivos de exclusión, se someterá al donante a un examen físico dirigido la toma de signos vitales y a revisar especialmente las zonas de punción. La piel de la zona de venopunción en ambos brazos no debe presentar lesiones o signos que indiquen que es adicto a drogas endovenosas. Lesiones poco importantes de la zona, como dermatitis y sarna serán causa de exclusión temporal. 4.- CONDICIONES DE LA ENTREVISTA DEL DONANTE DE SANGRE
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
5.- PRINCIPIOS DE UNA ENTREVISTA: 1) Iniciar -CEOR: Cállese, escuche, observe, registre. En general a las personas les resulta interesante hablar de sí mismos, por lo tanto al aplicar este principio se consigue más información. Este principio permite que el donante le hable, se explaye. Es importante observar su comunicación no verbal (gestos, posiciones, miradas, movimientos). Escuchar activamente le permitirá captar el mensaje, detectar incongruencias o nerviosismo, que pueden ser una pista para obtener mayor información. Puede obtener información a nivel consciente o subconsciente; esta última se conoce con el nombre de tincadas, corazonada, palpito, etc. Debe sondear y luego adquirir más información, contrastar para así decidir si solo es una “idea” o realmente es un antecedente de importancia. Debe hacer hablar al entrevistado por medio de preguntas abiertas.
2)Mantener el control de la entrevista: Con una actitud serena, objetiva, clara, conociendo la estructura de la entrevista, no hay posibilidad de que el entrevistado maneje la situación. Frente a un donante agresivo, prepotente, seductor, coqueto, amistoso, lloroso, angustiado, etc. lo importante es que el entrevistador (usted) no se involucre; hay que observar su actuación, ya que todas estas manifestaciones son en su mayoría mecanismos de defensa. Un donante agresivo, prepotente o expansivo es más fácil de conocer y de obtener información, ya que basta con dejarlo hablar. Por el contrario, si el donante es evasivo debe confrontarlo ya sea en forma directa o indirecta (ingenuamente). Frente a un donante alterado emocionalmente, debe darle tiempo a que se tranquilice pero no involucrarse en su emoción.
3)Escuchar activo: Significa leer las características del donante y concentrarse en el foco externo, observando cuidadosamente su conducta no verbal. verbal. Recordar los temas a tratar y así organizar el orden de la entrevista. 4)Tolerancia: Es la “aceptación de lo que sea” dentro del ámbito de la entrevista. No corresponde
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
6)Uso del silencio: Es fundamental para escuchar. Debe hacer un contacto visual laxo, permanecer en silencio con semi sonrisa, ladear ligeramente la cabeza para intensificar el efecto ya que es una invitación a hablar. Si el donante deja de hablar, debe mantener la posición de silencio, no interrumpir. Esto hace que el donante se incomode, sube su ansiedad, y trata de romper el silencio, por lo tanto hablará y entregará información. Los silencios del donante frente a una pregunta por ejemplo de su vida sexual, puede significar que él está evaluando su respuesta. Es importante estar alerta a los silencios del donante y debe volver a sondear, preguntar y confrontar. Es importante que recuerde, el silencio del donante de sangre no significa necesariamente que terminó de hablar, sobre todo si la información que le entregó le merece duda, sino que está organizando el pensamiento.
7)Facilitación: Si realiza pequeñas intervenciones pueden animar al donante para que hable o amplíe un tema en particular. Por ejemplo: Verbal: Sí, ya, ya veo, hum, ajá. Preguntas cortas: ¿Cómo así? “Ud. mencionó”. Repetir la última frase que dijo el donante (con el fin de que continúe el relato), resumir en una frase lo que dijo (técnica del reflejo – sirve para que amplíe el tema). No verbal: cabeza, sonrisa, levantar cejas, levantar hombros. Cuide especialmente las facilitaciones no verbales, debe administrarlas en momentos muy precisos ya que pueden llevar implícitamente a una valoración del tema que puede inhibir o ampliar la respuesta del donante, recogiendo errores en la respuesta; es recomendable que no las utilice. Recuerde que durante la entrevista debe mantener una expresión neutra que no induzca la respuesta del donante. En resumen debe facilitar que el donante hable pero mantener una expresión neutra.
8) Confrontación y contrastación: Contrastar durante toda la entrevista las inconsistencias y dudas que le merecen las respuestas del donante, para confrontárselas al final y así obtener respuestas más fidedignas. La contrastación es un ejercicio que debe realizar en forma íntima, señalando con asteriscos o marcas aquellos aspectos que ameritan ser confrontados. Es necesario confrontar hasta quedar tranquilo, pero cuidando que no sea molesto o
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
6.- CRITERIOS DE SELECCIÓN (Basados en la Norma General Técnica Nº 0146 – 2013: Norma que regula el procedimiento de atención de donantes de sangre (en sitio fijo o móvil). 1.- EDAD: - entre 18 y 60 años. Los donantes regulares que han donado una vez al año en los últimos 5 años se pueden aceptar hasta los 65 años, si cumplen con los requisitos de la entrevista y exámenes pre donación. 2.- FRECUENCIA DE DONACIÓN: debe existir un intervalo mínimo entre donaciones de sangre de 3 meses en los hombres y 4 meses en las mujeres. 3.- PESO: Mínimo 50 kilos. ESTATURA : Verificar que esté relacionada con el peso. Los donantes que pesan 50 Kgs. o más, pueden donar un máximo de 450 ml +/- 10% de sangre, es importante que las muestras para la calificación biológica de la donación, no excedan los 30 ml. La obtención de más del 13% de la volemia en un donante de 50 Kgs. puede significar una mayor frecuencia de reacciones adversas en el donante. El volumen debe ser controlado para asegurar que los componentes sanguíneos obtenidos cumplan con las especificaciones establecidas. 4.- ALIMENTACIÓN : los donantes que han comido en los horarios habituales, considerando cuatro ingestas alimentarias por día, pueden donar. Si un donante se ha saltado una comida se le debe dar a beber algo de líquido antes de la extracción. Se recomienda la ingestión de 500 mL de agua antes de la donación, ya que reduce significativamente la incidencia de reacciones adversas. 5.- PRESION ARTERIAL: (Tomada en reposo) - Sistólica: menor a 180 mmHg - Diastólica: menor a 100 mmHg 6.- PULSO: Entre 60 y 100 pulsaciones por minuto, regular y palpado por 30 segundos a lo menos. 7.- HEMOGLOBINA:sobre 13,5 grs/dL en hombres sobre 12,5 grs/dL en mujeres
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
! ! ! ! ! ! ! ! ! !
Examen físico Consentimiento informado Firma del profesional y del donante Antecedentes de la extracción Características de la bolsa Incidentes de la venopunción Identificación del técnico paramédico Reacciones adversas Tipo de donante Lugar de colecta
Cuestionario Preguntas ! Responsabilidad del donante sobre veracidad de las respuestas ! Aprobación – exclusión !
ALGUNAS PREGUNTAS A REALIZAR DURANTE LA ENTREVISTA AL DONANTE !
¿HA LEÍDO Y COMPRENDIDOEL INFORMATIVO? INFORMATIVO? Dirigida a saber si recibió, leyó y comprendió los folletos explicativos que contienen los requisitos generales establecidos para donar sangre,para protegerlo como donante y para proteger al paciente/ receptor de ella.
!
¿DURMIÓ MÍNIMO 5 HORAS?
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
Dirigida a detectar alteraciones o problemas en donaciones previas. Debe quedar registrada la característica de la reacción adversa a la donación.
SI: No debe donar si tiene historia de un desmayo severo o 2 desmayos consecutivos. si se decide aceptar, se requiere una observación cuidadosa. A CRITERIO: Se acepta como donante. NO: INFORMACION ADICIONAL: Una historia de predisposición a desmayos aumenta las posibilidades de una reacción adversa a la donación Debe excluirse a las personas que en donaciones anteriores, tuvieron un desmayo severo o en donaciones sucesivas tuvieron desmayos. !
¿HA SIDO RECHAZADO RECHAZADO ALGUNA VEZ COMO DONANTE DE SANGRE? ¿POR QUÉ? Dirigida a conocer alguna causa temporal o definitiva que lo inhabilite como donante. Enfatizar acerca del conocimiento de resultados alterados de exámenes realizados a su sangre. Excluir todo donante con antecedentes de exámenes positivos para: Hepatitis, Chagas, Sida, HTLV-I. En caso de Sífilis dependerá si hubo tratamiento, aceptándolo si existió y fue dado de alta hace un año por lo menos. Evaluar si el rechazo anterior fue por una causa definitiva o transitoria, en este último caso, el donante se acepta si el plazo determinado se ha cumplido. SI: NO:
!
Preguntar la causa y verificar si es causal o no de rechazo. Se acepta como donante.
¿HA TOMADO ASPIRINA O ANTINFLAMATORIOS EN LOS ÚLTIMOS 3 DÍAS? La sangre de donantes que hayan ingeridoaspirina o piroxicam los últimos 3 días o ha
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
OBLIGATORIO: no debe donar si han transcurrido menos de 8 semanas de la vacunación con microorganismos vivos: • • • • • •
Fiebre amarilla Parotiditis Polio oral Rubeola Sarampión Tifoidea oral
A CRITERO: aceptar si la vacunación se realiza con microorganismos no vivos: Ántrax Botulismo Cólera Difteria Haemophilus influenzae Influenza Meningococo Neumococo Polio inyectable Tifoidea inyectable Coqueluche • • • • • • • • • • •
CASOS ESPECIALES: consultar NGT 146 para mayores detalles: Rabia: si se administró después de una mordedura animal, no debe donar hasta 12
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
• • • • • • •
La cirugía se debió a un cáncer No han sanado todas las heridas Hay cualquier infección No se ha restablecido la movilidad normal Han transcurrido menos de 12 meses de una cirugía mayor Han transcurrido menos de 7 días de cualquier otra cirugía Si requiere tratamiento post operatorio o atención hospitalaria regular
NO: !
Acepte al donante
¿EN LOS ÚLTIMOS 12 MESES SE HA REALIZADO ENDOSCOPIA? El antecedente de endoscopía, está destinado a detectar donantes en riesgo de infección por hepatitis B o similares, con el uso de endoscopio flexible.
NO: SI: ! •
!
Se acepta como donante Evaluar según norma
¿SE HA REALIZADO EXAMEN DE SANGRE U OTRO TIPO DE EXAMEN? OBLIGATORIO: no debe donar si: Está en espera de exámenes o de resultados para una condición no diagnosticada aún y que podría significar el rechazo de la donación. ¿EN LOS ÚLTIMOS 6 MESES HA RECIBIDO ALGÚN TRATAMIENTO MÉDICO? ¿HA TOMADO ALGÚN MEDICAMENTO EN EL ÚLTIMO TIEMPO? ¿CUÁL?
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
! !
¿EN LOS ÚLTIMOS 12 MESES HA IDO AL MÉDICO, MATRONA, DENTISTA U OTRO PROFESIONAL DE SALUD?
!
¿HA TENIDO CÁNCER ALGUNA VEZ?
!
¿TIENE O HA TENIDO ALGUNA ENFERMEDAD AL CORAZÓN, PULMÓN, RIÑON, TIROIDE O TIENE HIPERTENSIÓN, DIABETES, ALERGIA, TENDENCIA A SANGRAR U OTRA?
Preguntas dirigidas a detectar patologías que lo inhabiliten como donante. a) Enfermedades Cardiovasculares: Se excluyen: Cardiopatías con insuficiencia cardíaca, Enfermedad reumática en tratamiento y Enfermedad coronaria (angina, infarto), por la posibilidad de una nueva crisis. b) Diabetes: Se excluye quienes están bajo tratamiento con medicamentos normoglicemiantes, insulinodependientes. Se aceptan los que controlan su diabetes sólo con régimen. c) Hipertensión: Se excluye aquellos que estén en tratamiento. En caso de detectar valores de presión fuera de los rangos establecidos, se debe esperar 15 minutos y volver a tomar la presión con el donante acostado. d) Enfermedades Gastrointestinales: Se excluye las diarreas agudas o crónicas reactivadas. Se aceptan donantes con antecedentes de úlcera péptica asintomáticos y sin tratamientos. Otras patologías como pólipos rectales o hemorroides, pueden ser aceptados como donantes si en el momento del interrogatorio están asintomáticos y sin tratamiento médico.
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
o) Sífilis y Gonorrea: Se excluye por un período de 12 meses, ya que pudieran asociarse con transmisión de VIH. p) Sangramientos importantes: Consultar causa, frecuencia y temporalidad. Se excluye si es frecuente, importante y activo. q) Enfermedades crónicas: Dirigida a detectar patologías que lo inhabiliten como donante. En caso de exclusión permanente, explicarle que nunca debe donar sangre y por qué. r) Pérdida inexplicable de peso: El peso es el parámetro biomédico de gran importancia. En el adulto tiende a mantenerse estable en el tiempo. La ganancia de peso por sobre los valores normales para estatura y sexo se definen como obesidad. La pérdida injustificada de peso (4 - 5 Kg. y más en los últimos meses sin causa aparente) puede constituir un elemento de juicio médico indicador de enfermedad crónica como: cáncer, SIDA, TBC, hipertiroidismo, anorexia psicógena, etc. Por lo tanto, constituye motivo de rechazo como donante de sangre. Hace excepción la baja de peso motivada por régimen, asistida por médico. !
¿EN LOS ÚLTIMOS 12 MESES LE HAN PUESTO SANGRE A USTED O A SUPAREJA SEXUAL?
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
documenta una hepatitis de tipo A, puede ser aceptado como donante, si han pasado más de 12 meses de su recuperación. !
¿HA RECIBIDO TRATAMIENTO CON HORMONA DE CRECIMIENTO ANTES DE 1985? No debe donar si ha recibido gonadotrofinahipofisaria u hormona de crecimiento de origen humano.
!
¿HA VIVIDO EN INGLATERRA ENTRE LOS AÑOS 1980 Y 1996 O HA RECIBIDO ALLÍ TRANSFUSIÓN DE ALGÚN COMPONENTE SANGUÍNEO?
!
¿TIENE HISTORIA FAMILIAR DE LA ENFERMEDAD DE CREUTZFELDT JAKOB? Estas preguntas están enfocadas a prevenir la transmisión de la enfermedad de Creutzfeldt-Jakob, una alteración rara, degenerativa y fatal del sistema nervioso. No se ha
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
!
EN LOS ÚLTIMOS 12 MESES, ¿UD. O SU PAREJA SE HAN REALIZADO ALGÚN TATUAJE, PIERCING, ACUPUNTURA, O HA SUFRIDO ALGÚN PINCHAZO ACCIDENTAL CON AGUJA O JERINGA CON SANGRE? Dirigida también a la identificación de personas que pueden estar en fase de incubación de VIH, hepatitis u otra infección de transmisión sanguínea. Personal de salud no se considera grupo de riesgo, a menos que haya sufrido accidentes corto-punzantes con material presuntamente contaminado, en los últimos 12 meses, en cuyo caso debe excluirse por igual periodo. NO: Se acepta al donante. Si: Tatuajes: se excluye por un periodo de 12 meses Acupuntura y perforación auricular: se excluye por un periodo de 12 meses NOTA: preguntar por tratamientos inyectables con material no desechable, ya que sería causal de exclusión.
!
¿HA COMPARTIDO AGUJA O JERINGA CON OTRA PERSONA?
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
Preguntas dirigidas a detectar personas que están en riesgo de infección por VIH y otras enfermedades infecciosas que podrían ser transmitidas al receptor de productos sanguíneos.
NO: Se acepta como donante. SI: Evaluar el riesgo real y la promiscuidad de la conducta sexual. Si es el caso, excluir por 12 meses !
¿HA TENIDO CONTACTO SEXUAL CON ENFERMO O PORTADOR CRÓNICO DE HEPATITIS B o CEN LOS ÚLTIMOS 12 MESES? Dirigida a identificar pacientes que pueden estar en fase de incubación de Hepatitis. Consultar Norma. NO: SI:
Se acepta Donantes con antecedentes de este tipo de contacto se excluyen por 12 meses.
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
PROCEDIMIENTOS A UTILIZAR EN LA ATENCION Y SELECCION DE UN DONANTE DE SANGRE I.- DETERMINACION DE Hb UTILIZANDO SULFATO DE COBRE. PRINCIPIO: Se basa en la gravedad específica. Una gota de sangre introducida en una solución de Sulfato de Cobre queda encapsulada por proteinato de cobre, que evita cualquier cambio de gravedad específica durante 15 segundos. Si la gota tiene una gravedad específica superior a la solución, caerá en 15 segundos. Si no es así, la gota oscilará, permanecerá suspendida o subirá hacia la superficie de la solución. Es una prueba cualitativa que demuestra sólo si la hemoglobina se encuentra por encima o por debajo de límites aceptables. MUESTRA: Sangre, obtenida por punción capilar del pulpejo del dedo, de acuerdo a
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
4.-
Deje caer suavemente del tubo capilar 1 gota de sangre desde una altura de aproximadamente 1 cm. sobre la superficie de la solución de Sulfato de Cobre.
5.-
Observe durante 15 segundos. Si la gota de sangre se va al fondo, el nivel de Hb.
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
3. 4. 5. 6. 7.
Microcubetas HemoCue para determinación Lancetas desechables Algodón Desinfectante Caja de descarte de material cortopunzante
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
7. Tomar la cubeta por la parte posterior y poner en contacto con la sangre la porción donde se encuentran los reactivos de manera de llenarla completamente por capilaridad. Evite contaminar el ojo óptico. No rellene la cubeta. Si hay burbujas de aire en el ojo óptico de la cubeta debido a un lleno inadecuado de la cubeta, debe descartarla y utilizar una nueva para realizar el test.
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________________________________________________________ ___________________________ ________________________________________________________INMUNOHEMATOLOGIA ___________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
- Baguetas PROCEDIMIENTO. 1.-Coloque en una lámina limpia y seca una gota de cada uno de los antisueros, primero
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_______________________________________________________ __________________________ __________________________________________________________ _____________________________INMUNOHEMATOLOGIA INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
IV.- CONTROL DE CALIDAD DEL PROCEDIMIENTO PROCEDIMIENTO DE EXTRACCION EN UN DONANTE DE SANGRE. 1.- CULTIVO BACTERIOLÓGICO DEL SITIO DE PUNCIÓN. Debe realizarlo una vez al mes o según lo establezca el manual de procedimientos del
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
Reacción de células con Reacción del suero con células Interpretación antisueros testigo. GRUPO Anti-A Anti-B Células A Células B 0 0 + + O + 0 0 + A 0 + + 0 B + + 0 0 AB 0= no aglutinación += aglutinación Cuando los resultados entre ambas pruebas no concuerdan, existe una discrepancia, la que debe ser investigada antes de informar el grupo ABO. Si la discrepancia se presenta en una unidad de sangre donada, ésta debe mantenerse aparte y no transfundirla hasta resolver el problema. Cuando la discrepancia se presenta en un paciente que requiere una transfusión, él puede ser transfundido con eritrocitos concentrados del grupo O, Rh compatible, para solucionar la emergencia, teniendo la precaución de obtener previamente una muestra suficiente como para resolver el caso. Las discrepancias en una clasificación ABO pueden deberse a:
1- Errores técnicos - Material sucio. - Contaminación bacteriana de las muestras. - Contaminación de reactivos, sueros y células. - Proporción incorrecta de la relación suero/células - Exceso o falta de centrifugación - Falta de adición de reactivos y muestra - Temperatura de incubación incorrecta - Pérdida de actividad de los reactivos - Empleo inadecuado de reactivos y células testigos - Presencia de hemólisis en la muestra - Identificación incorrecta de la muestra, tubos. - Registros mal llevados e incompletos. - Etc. 2- Problemas inherentes a las células - Presencia de gelatina de Wharton en muestras de cordón - Pacientes recientemente transfundidos. -Autosensibilización de los GR del paciente (autoanticuerpos) o EHRN - Poliaglutinación - Antígenos débiles de A y/o B -Pacientes que sufren de patologías infecciosas importantes (Ag. B adquirido) u otras patologías malignas. -Presencia de altas concentraciones de sustancias de grupo sanguíneo en el plasma. - Etc.
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
- Etc. CONTROL DE CALIDAD: a.-Tipificación de antígenos: GR A2, A2B, B, O y autocontrol. b.-Detección de anticuerpos: Suero AB inerte c.-Chequear los GR. testigos
Estos controles deben incluirse en cada serie de tests. PREGUNTAS A RESPONDER: 1.- ¿Cuál es la causa de cada uno de los factores de error descritos? 2.- ¿Cómo interfieren en la clasificación ABO los factores de error descritos? 3.- ¿Producen resultados falsos positivo o negativo? 4.- ¿Existen otras causas de error en la clasificación ABO que no aparezcan en este listado? 5.- ¿Cómo se resuelve cada una de las discrepancias indicadas? 6.- Elabore un esquema a aplicar en el laboratorio en que se describa paso a paso lo que debe realizarse en la resolución de una discrepancia. Existen subgrupos de A y B, que corresponden a expresiones fenotípicas más débiles de los antígenos habituales y que en el laboratorio pueden investigarse apoyándose en el siguiente esquema:
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
SUBGRUPO
A1 Aint A2 A3 Ax Ael Am B B3 Bx
REACCION CON ANTISUEROS Anti Anti Anti Anti A AB A1 H 4+ 4+ 4+ 0 4+ 4+ 2+ 3+ 4+ 4+ 0 2+ 2+# 2+# 0 3+ 0/º 2+ 0 3+ 0 0 0 4+ 0/º 0/º 0 4+ 0 4+ 0 0 0 2+# 0 4+ 0 0/2+ 0 4+
*: ocasionalmente forman anti-A 1
REACCION CON ERITROCITOS A1 A2 B O 0 0 0 0 0* 0 0* 0 2+º 0 2+º 0 0 0 4+ 4+ 4+ 4+ 4+ 4+
4+ 4+ 4+ 4+ 4+ 4+ 4+ 0 0 0
0 0 0 0 0 0 0 0 0 0
ANTIGENOS EN SECRECION SALIVA AyH AyH AyH AyH H H AyH ByH ByH H
#: campo mixto. º: frecuentemente forman anti-A1.
TECNICA DE AGLUTINACIÓN EN COLUMNA ( TARJETA): Para realizar una determinación ABO con esta técnica, debemos utilizar una tarjeta que contenga columnas con Anticuerpos anti-A, anti-B, y 2 columnas neutras para realizar la preuba inversar con suero o plasma del paciente y glóbulos rojos testigos A1 y B. MUESTRA -Sangre completa con anticoagulante. MATERIALES -Pipeta automática. -Puntas desechables para pipeta. -Tubos Khan. -Gradilla para tubos Khan. -Centrifuga para tarjetas. REACTIVOS -Tarjetas de gel para tipificación ABO. -Solución salina. -Eritrocitos testigos anti- A1y anti- B. METODO -Numere las muestras a clasificar y colocarlas ordenadamente en una gradilla apropiada. -Colocar tras cada muestra un tubo khan numerado igual que la muestra. -Preparar en el tubo Khan una suspensión de hematíesen medio Salino(según instrucciones del fabricante). -Añadir la suspensiónde glóbulos rojos previamente hecha a las columnas identificadas como A, B y Ctl. - Añadir los GR testigos A1 a la columna identificada como N A1 y los GR testigos B a la
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
INTERPRETACIÓN La interpretación de la determinación de grupo ABO al igual que en tubo se realiza en cruces de 1 a 4.
DETERMINACION DE ANTIGENOS SOLUBLES ABH Y LEWIS EN SALIVA En Chile aproximadamente el 83% de la población presenta antígenos solubles del sistema ABO en la saliva, y por lo tanto poseen el gen Se, quien es el responsable de su presencia en este y otros fluidos corporales. RECOLECCION DE LA SALIVA 1.- Obtenga entre 5 a 10 ml de saliva en un tubo ancho, de preferencia enjuagándose la boca previamente con abundante agua. La mayoría de los individuos puede acumular esta cantidad en varios minutos. Para fomentar la salivación, el paciente puede mascar cera especial, lámina de parafina o cinta de goma, pero no chicle ni cualquier material que contenga azúcar o proteínas. 2.- Centrifugue la saliva a alta velocidad durante 10 minutos. Transfiera el sobrenadante a un tubo limpio. 3.- Coloque el tubo con la saliva en un baño María hirviendo por 10 min para inactivar las enzimas salivares. 4.- Obtenga en otro tubo el sobrenadante claro y transparente, descartando la parte sólida y opaca. Diluya la saliva con un volumen igual de salino. 5.- Refrigere si el test no se realizará en las próximas horas. Si este no se realiza en el día de colección, congele la muestra la que permanecerá estable por varios años. REACTIVOS: -(seleccione el reactivo de acuerdo al antígeno que desee detectar) -Anti-A y anti-B policlonal (humano). -Lectina anti-H de Ulex europeus. -Anti-Lea policlonal (conejo/cabra). -GR testigos A1 y B, de iguales características a los utilizados en la clasificación ABO. -GR grupo O, Lea +, con las características de los usados en detección de anticuerpos irregulares PROCEDIMIENTO: A.- Selección de la dilución del antisuero: 1.- Prepare diluciones seriadas del anticuerpo seleccionado. Puede usar lectina anti-H para ABO y anti-Lea para el sistema Lewis. 2.- Coloque en tubos previamente rotulados con la dilución correspondiente una gota de dilución y una gota de la suspensión al 2-5 % en salino de los eritrocitos apropiados. 3.- Centrifugue y observe en busca de aglutinación macroscópica. 4.- Seleccione para el test la menor dilución que presente una aglutinación de 2+. B.- Estudio de la condición de secretor: 1.- Marque 4 tubos como: control+, control-, test y control de dilución. 2.- Agregue a cada uno de los tubos ya marcados, una gota del antisuero apropiado y previamente seleccionado de acuerdo al procedimiento descrito en A.
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
5.- Agregue una gota de suspensión al 3% en salino de eritrocitos que posean el antígeno para el antisuero que se está utilizando. 6.- Mezcle el contenido del tubo e incube 30 a 60 minutos a temperatura ambiente. 7.- Centrifugue el tiempo estandarizado para lectura en salino en su centrífuga, y observe los tubos en busca de aglutinación macroscópica. Registre los resultados.
INTERPRETACION DEL TEST DE SALIVA Compare sus resultados con la siguiente tabla: Saliva Test
Saliva Se
Saliva no SE
2+ -
-
2+ 2+
Control Dilución 2+ 2+
Interpretación No Secretor Secretor
1.- La aglutinación de los GR testigos en el tubo test indica ausencia del correspondiente Ag en la saliva. 2.- No aglutinación de los GR testigos en el tubo test indica presencia prese ncia del Ag correspondiente en la saliva. 3.- Ausencia de aglutinación en el tubo control dilución indica ind ica que el reactivo (Ac) empleado está demasiado diluido. Debe repetir el paso A de este procedimiento. NOTA: Este test puede ser adaptado para hacer mediciones semicuantitativas de actividad de grupo sanguíneo al estudiar diluciones seriadas en salino de la saliva. Mientras más alta sea la dilución necesaria para eliminar la actividad neutralizante, mayor cantidad de antígeno ABH soluble existe.
CLASIFICACION Rh (D): TÉCNICA EN TUBO MUESTRA: Sangre con o sin anticoagulante. MATERIAL: - Pipetas Pasteur - Tubos Khan (11x75) - Gradilla para tubos - Lector para inmunohematología - Baño termo regulable - Centrífuga para Inmunohematología. REACTIVOS: - Suero anti-D: Existen sueros de alto contenido proteico, químicamente modificados de bajo contenido proteico, o mezclas IgM + IgG de bajo contenido proteico. Debe seguir cuidadosamente las instrucciones de uso del fabricante y hacer los ajustes pertinentes.
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
METODO: 1.- Enumerar las muestras a tipificar. 2.- Lavar por lo menos una vez una alícuota de eritrocitos de la muestra con PBS, y preparar con ellos una suspensión al 2-4% en salino, suero AB inerte u otro. (Puede usar la misma suspensión preparada para la clasificación ABO). 3.- Depositar 1 gotas de suero anti-D en un tubo Khan previamente marcado D. 4.- Colocar una gota de suero Rh control en otro tubo marcado, o albúmina al 30% si usa reactivos con alto contenido proteico (como control negativo). 5.- Agregar una gota de la suspensión de eritrocitos preparada en el punto 2 a ambos tubos. 6.- Centrifugar el tiempo establecido para lectura en la estandarización de su centrífuga. 6.- Girar suavemente el tubo y observar si existe o no aglutinación macroscópica frente a una buena fuente de luz. 7.- Si el resultado es negativo, incubar por un mínimo de 15 minutos y un máximo de 1 hora a 37ºC. (de acuerdo con las instrucciones del fabricante). 8.- Proceder igual que en los puntos 6 y 7.
INTERPRETACIÓN: -
Si hay aglutinación = o > a 2+ en el tubo test, y no se observa en el control, la persona presenta el antígeno D en los eritrocitos, por lo tanto es D+ (Rh positiva). Si la aglutinación es < de 2+ o negativa en el tubo test, no debe interpretarse como Rh + sino que debe continuarse el estudio en busca de expresiones débiles o variantes del antígeno D, para definir así el grupo Rh. Si el tubo control Rh presenta aglutinación, no informar antes de verificar con otro tipo de reactivo el grupo Rh de la muestra.
Investigación de antígeno D débil: 9.- Incubar ambos tubos cuya lectura fue negativa por 15 a 30 minutos minutos a 37ºC. 10.- Centrifugar y leer igual que en los puntos 5 y 6. 11.- Si el resultado es positivo en el tubo test y negativo en el control, la persona es Rh positivo. Si la lectura es negativa proceder como sigue: 12.- Lavar los eritrocitos de ambos tubos con abundante PBS por 4 veces. 13.- Después del último lavado, eliminar tanto salino como sea posible dejando los eritrocitos resuspendidos en el salino residual adherido a las paredes. 14.- Agregar una gota de suero Antiglobulina Humana.
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
En aquellos casos en que se obtiene un resultado positivo en el punto 17, y el test AGH directo (que se usa para confirmar que el anticuerpo adherido a los eritrocitos en estudio corresponde al anti-D agregado y no a un anticuerpo previamente adherido), es negativo, se debe informar como Rh +. Si el AGH directo es positivo, no se puede informar Rh, si se trata de un receptor de transfusión debe usarse sangre Rh negativa mientras se estudia su Rh. Es incorrecto informar una muestra como Rh - Du +, la nomenclatura actual se refiere al Du como un antígeno débil pero Rh positivo, lo que implica presencia de él en los eritrocitos. El cuadro siguiente muestra algunos posibles resultados:
ANTI-D
CONTROL Rh
+ + -
+ -
INV. D DEBIL (Du) + +
TAD
INFORME
+
Rh positivo ¿ Rh negativo Rh positivo ¿
En aquellos casos en que no se puede informar el Rh (?), se debe utilizar otro tipo de reactivo Rh y eventualmente aplicar otras técnicas como elución y absorción.
CONTROL DE CALIDAD: Todos los resultados son válidos si se usan en cada serie de tests, controles adecuados usando células con antígenos débiles y negativos para el antígeno en estudio, ej.: GR R1r y rr para anti-D, y se validan los resultados de AGH negativos con células control de AGH. RESULTADOS FALSOS: Las causas de que se produzcan reacciones falsas en la determinación del grupo Rh, pueden asociarse a: 1.- Uso de reactivos de alto contenido proteico: Por su contenido pueden causar reacciones falsas positivas si los hematíes del paciente están recubiertos de anticuerpos. La incubación de los GR y el antisuero durante un tiempo excesivo, antes de efectuar la lectura de la prueba, puede tener como consecuencia que el reactivo con alto
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
4.- Hematíes poliaglutinables pueden ser aglutinados por cualquier reactivo que contenga suero humano, pero rara vez existe este error ya que la edad, dilución y los diferentes pasos del proceso de fabricación, tienen tendencia a eliminar estos anticuerpos. 5.- Contaminación bacteriana de los reactivos, o con sustancias extrañas, o con reactivos provenientes de otros frascos. Es recomendable nunca sacar a la vez el cuentagotas de más de un reactivo, y debe inspeccionarse periódicamente los frascos en busca de deterioro. Es importante recordar que en los reactivos de alto contenido proteico, la contaminación bacteriana puede no producir una turbidez detectable, ya que el índice de refracción de las bacterias es similar al del propio material. 6.- No se ha añadido el antisuero al tubo. Un buen habito es fomentar el colocar en primer lugar el antisuero al tubo, controlar su presencia y luego colocar la suspensión de hematíes. 7.- Un antisuero específico no reacciona con un antígeno variante o deprimidos. 8.- Es posible que un antisuero que contenga un anticuerpo dirigido principalmente contra un antígeno compuesto Rh no presente una reacción detectable con los hematíes portadores de antígenos individuales como producto de genes independientes. Esto ocurre más frecuentemente con los sueros anti-C, cuyas reacciones con el antígeno C, casi invariablemente son más fuertes cuando este ha sido producto de R1 o r'. 9.- No se siguen las instrucciones de los fabricantes y en consecuencia se ha usado incorrectamente el antisuero. 10.- Si se realiza una agitación demasiado enérgica al resuspender el botón de hematíes después de la centrifugación, puede dispersarse una aglutinación débil. 11.- Reactivo inactivo por falla en e n su manipulación y almacenamiento. El anti-D químicamente modificado es especialmente sensible a la destrucción del IgG por enzimas proteolíticas que podrían producir las bacterias.
TÉCNICA DE AGLUTINACION AGLUTINACION ENCOLUMNA MUESTRA Sangre completa con anticoagulante. MATERIAL Micropipetas Puntas para micropipetas Tubos khan. Gradilla para tubos khan. Incubador para tarjetas. Centrifuga para tarjetas
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
menor a 2+ deben ser confirmadas.
ALGORITMO A UTILIZAR EN LA CLASIFICACIÓN Rh D MUESTRA SANGRE
con o sin anticoagulante
Suspensión eritrocitos eritrocitos al 3% en PBS
GR 3% + anti-D
POSITIVO Controles Controles OK OK
NEGATIVO
INFORME Rh Positivo
AGH DE Rh o test Du
NEGATIVO Controles OK
POSITIVO
INFORME Rh negativo
NO INFORMAR
REALIZAR TAD
TAD NEGATIVO
TAD POSITIVO
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
CLASIFICACION CLASIFICACION DE GRUPO ABO Y Rh: TÉCNICA EN MICROPLACA Las técnicas en microplacas pueden usarse para investigar antígenos ABO en los glóbulos rojos y anticuerpos en el suero. La microplaca es una matriz de 96 tubos de ensayo, en los cuales puede visualizarse hemaglutinación con sueros clasificadores diluidos y en bajo volumen. Las microplacas pueden ser flexibles o rígidas, con microcubetas en forma de U o V. Las más usadas son las en forma de U porque permiten la lectura de los resultados después de centrifugar la placa y observar las características de los GR resuspendidos o analizar el patrón de flujo de las células cuando se inclina la placa, pudiendo finalmente estimar la intensidad de la aglutinación. MUESTRAS: -Adultos: sangre con EDTA -Recién nacidos: sangre de cordón. MATERIALES: -Microplacas con fondo en “U” -Pipetas Pasteur. -Tubos de Kahn. -Gradillas. EQUIPOS: -Centrífugas con soporte para microplacas. -Agitador. -Lector. REACTIVOS: -Sueros clasificadores anti-A y anti-B monoclonales diluidos, según estandarización hecha previamente. -Suero clasificador anti-D, diluido según estandarización. -Glóbulos rojos A-1,A-2, B, O R1r y O rr suspendidos al 1% en EDTA. -PBS pH 7.3. TÉCNICA:
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
1. Repartir los sueros de la siguiente forma: una gota de anti-A en todos los pocillos de la primera fila, una gota de anti-B en la segunda fila , una gota de anti-AB en la tercera fila, una gota de anti -D en la cuarta y quinta fila y una gota de PBS en la sexta fila. 2. Repartir las muestras de la siguiente manera: una gota de suero en los pocillos G y H. 3. En un tubo aparte prepare una suspensión de GR de la muestra a estudiar al 1% en PBS, reparta una gota de esta suspensión en los pocillos A al F. 4. Reparta una gota de la suspensión de GR A1 en los 10 primeros pocillos de la fila G. 5. Reparta una gota de la suspensión de GR B en la fila H. 6. Reparta 1gota de las suspensiones de GR control según se muestra en el esquema anterior. 7. Centrifugue las placas a 1400 r.p.m. durante 20 segundos. 8. Agite las placas durante 20 seg. Con el agitador en alta revolución y 20 seg. Con el agitador en baja revolución. 9. Lleve las placas al lector para ver presencia o no de aglutinación. Registre el resultado en el protocolo correspondiente en intensidad de cruces, siguiendo el mismo criterio que en la clasificación en tubo. Si existe alguna discrepancia en la clasificación se debe repetir en tubo. Del mismo modo, todas las muestras que resulten negativas con anti-D, se deben clasificar en tubo. ABO Rh
B A A A O O A O O B AB A + + + + + + + + + + + +
DETERMINACION DETERMINACION DE GENOTIPO RH
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
Para realizar este cálculo debemos recurrir a la frecuencia de los haplotipos Rh en la población chilena y basarnos en el equilibrio de Hardy-Weinberg. Si se desea calcular por ejemplo la frecuencia en Chile del genotipo CDe/cde, debe calcularse 2 X (CDe)(cde) = 2 (0.500 X 0.2192) = 0.22
Frecuencia de los haplotipos Rh en población chilena. CDE (Rz) CDe (R1) cDE (R2) cDe (Ro) CdE (Ry) Cde (r') cdE (r'') cde (r)
0.0132 0.5001 0.2259 0.0255 0.0000 0.0064 0.0097 0.2192
REFERENCIA: Rev. Med. de Chile 116: 28-33. 1988.
NUMERO DE SITIOS ANTIGENICOS DE ACUERDO AL GENOTIPO CELULAR GENOTIPO R1R1 R1r R1R2
D 14.500 19.300 9.900 14.600 23.000 31.000
C 46.000 56.000 21.000 40.000 25.000 40.000
ANTIGENO E 15.000
c 37.000 53.000 37.000 53.000
e 18.200 24.000 18.200 24.000 13.400 14.500
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
FRECUENCIAS GENICAS DE SISTEMAS SANGUINEOS EN DIFERENTES POBLACIONES SISTEMA
MIXTA CHILENA
ESPAÑOLA
MAPUCHES AYMARAE ATACAMEÑ S OS
NEGRA
ABO A B O N:
0.1824 0.0698 0.7477 321
0.2864 0.0670 0.6465 20000
0.018 0.000 0.9820 148
0.0549 0.0160 0.9286 183
0.0562 0.0027 0.9407 182
0.1780 0.1143 0.7077 858
Duffy Fya No Fya N:
0.5805 0.4195 379
0.3966 0.6034 1988
0.7080 0.2920 141
0.7823 0.2177 654
0.7008 0.2990 180
0.0607 0.9393 365
Rh CDE CDe .cDE .cDe CdE Cde .cdE .cde
0.0093 0.4999 0.2224 0.0230 0.0000 0.0089 0.0139 0.2223
0.0008 0.4036 0.1670 0.0186 0.0000 0.0049 0.0029 0.3820
0.0320 0.6930 0.2550 0.0210 0.0000 0.0000 0.0000 0.0000
0.0267 0.3657 0.5757 0.0329 0.0000 0.0000 0.0000 0.0000
0.0465 0.4727 0.4340 0.0267 0.0000 0.0000 0.0000 0.0198
0.0000 0.0256 0.0427 0.7395 0.0000 0.0707 0.0000 0.1184
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
TEST ANTIGLOGULINA HUMANA
-
El test Antiglobulina Humana (AGH) o test de Coombs, se basa en principios elementales simples: Las moléculas de anticuerpos y los componentes del complemento son globulinas. Al inyectar a un animal globulina humana, se estimula la producción de anticuerpos frente a esta proteína extraña. Se remueven del suero las proteínas y anticuerpos no deseados dejando solamente el ANTICUERPO ANTIGLOBULINA HUMANA. Este suero AGH reaccionará específicamente con globulinas humanas tanto si están unidas a eritrocitos como libres en el suero. La porción Fab de la molécula AGH reaccionará con la porción Fc del anticuerpo unido a cada glóbulo rojo por separado produciendo la aglutinación. Las globulinas libres no unidas, pueden neutralizar el AGH si no se eliminan completamente y causar resultados negativos falsos. Es uno de los test de laboratorio más usados en inmunohematología, se utiliza 2 tipos de test AGH 1. 2.
- El test Antiglobulina Directo TAD. TA D. - El test Antiglobulina Indirecto TAI.
TEST ANTIGLOBULINA HUMANA DIRECTO (TAD) TEST DE COOMBS DIRECTO TÉCNICA EN TUBO El TAD se utiliza para demostrar anticuerpos IgG y/o complemento (C3dg), que se han unido "in vivo" a los eritrocitos del paciente. Se utilizan directamente los eritrocitos lavados del
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
4.- Si al usar suero sue ro AGH poliespecífico poliespecífic o o anti-C3d la reacción reac ción es negativa, incube incu be por 5 minutos a temperatura ambiente el tubo y proceda igual que en el punto 3. Esto le otorga mayor sensibilidad a la detección del complemento, especialmente en anemias hemolíticas autoinmune. Esta segunda lectura jamás puede reemplazar a la lectura inmediata ya que la reacción del anticuerpo IgG puede hacerse débil o negativa. 5.- Agregue 1 gota de células control de AGH (GR sensibilizados con IgG) a todo test cuyo resultado es negativo. 6.- Proceda igual que en el punto 3. Si los eritrocitos de d e la muestra no fueron adecuadamente lavados, quedando Igs libres, el suero AGH se neutralizará obteniendo en esta fase un resultado negativo. Para que el resultado del TAD sea válido, el resultado de esta etapa debe ser positivo. INTERPRETACIÓN: 1.- El TAD es positivo cuando se detecta aglutinación desde la centrifugación inmediata o después de la incubación de 5 minutos a temperatura ambiente. Las reacciones por IgG se observan en la centrifugación inmediata y el complemento generalmente se presenta mejor luego de la incubación a temperatura ambiente. Es necesario el uso de SAGH monoespecíficos para confirmar el tipo de globulinas presentes. 2.- El TAD es negativo cuando no se observa aglutinación en la centrifugación inmediata, luego de la incubación a temperatura ambiente, y si aparece la reacción positiva al agregar los eritrocitos sensibilizados con IgG en la última fase. NOTA: Un test con resultado negativo no necesariamente indica ausencia de globulinas sensibilizando los eritrocitos, los sueros AGH poliespecíficos y anti-IgG, detectan aproximadamente 200-500 moléculas de IgG por eritrocito, se han publicado casos de AHAI con un número inferior de anticuerpos por célula y por lo tanto con un TAD negativo.
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
TEST ANTIGLOBULINA HUMANA INDIRECTO (TAI) TEST DE COOMBS INDIRECTO TÉCNICA EN TUBO El Test Antiglobulina Indirecto (TAI) se utiliza para comprobar el recubrimiento "in vitro" de los eritrocitos por anticuerpos y/o complemento, mediante incubación del suero (Ac) con hematíes (Ag) que posteriormente se lavan para eliminar las globulinas no adheridas. El hecho de que se produzca aglutinación después de añadir el reactivo AGH indica que el suero contiene anticuerpo(s) que reacciona(n) con antígeno (Ag) presente en el glóbulo rojo. Se utiliza para detección, identificación y titulación de anticuerpos, pruebas cruzadas, determinación de grupos sanguíneos con ciertos antisueros. TAI USANDO GLOBULOS ROJOS SUSPENDIDOS EN SALINO
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
12. Leer en busca de hemólisis y/o aglutinación, rotando suavemente el tubo frente a una fuente de luz. Anotar los resultados tubo en mano. 13. Confirmar los resultados negativos, agregando 1 gota de células control AGH (Coombs) a los tubos en que el resultado del test fue negativo. 14. Centrifugar y leer según punto 4. INTERPRETACIÓN: Se considera que un TAI es positivo cuando se detecta aglutinación después de agregar AHG. Esta positividad significa presencia de anticuerpo(s) de tipo IgG y/o de anticuerpo(s) que se manifiesta a través de la fijación de Complemento. Para que el test se considere válido, el tubo con suero control positivo debe presentar un resultado positivo y el resultado de las células control AGH debe ser positivo. NOTA: Puede acortarse el tiempo de incubación si en el punto 3, los eritrocitos se suspenden en medio de baja fuerza iónica (LISS), teniendo la precaución de utilizar los tiempos y volúmenes previamente estandarizados para este reactivo.
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
FASE 1: Selección de los eritrocitos de prueba: 1.- Deben ser grupo O Rh positivo y contener todos todos los antígenos de importancia clínica. 2.- Usarse por separado, nunca en pool. 3.- Prepararse una suspensión al 2-4% en PBS, y 1,5 - 2% en LISS FASE 2: Sensibilización de los eritrocitos: 1.- Proporción correcta suero/células: suero/células : para TAI en salino = 2:1 a 4:1 para TAI en LISS = 1:1 2.- Tiempo de incubación: Para TAI salino = 45 minutos a 1 hora Para TAI LISS = 15 a 20 minutos, según la estandarización del test. FASE 3: Lavado de las células: 1.- Trabajar con centrífugas calibradas. 2.- Uso de 4 lavados con abundante PBS. 3.- No interrumpir el proceso de lavado en ningún momento ya que los anticuerpos pueden eluirse.
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
- Eritrocitos grupo O Rh positivo (pool de 4 células R1r, con TAD negativo y con menos de 3 semanas de almacenamiento) - Anti-D con título 32 o 64. METODO: 1.- Lave los eritrocitos seleccionados por 3 veces con abundante PBS. 2.- Coloque 10 ml de PBS en un tubo y agregue 10 gotas del suero anti-D. Agregue una cantidad suficiente de los eritrocitos O previamente lavados como para obtener una concentración al 3% (aprox. 150 microlitros) 3.- Mezcle e incube a 37ºC durante 30-60 minutos. 4.-Lave los glóbulos rojos 4 veces con PBS y resuspendalos hasta alcanzar una concentración final del 2-3%. 5.- Realice un TAD a los eritrocitos tratados con el suero Antiglobulina Humana que Ud. utiliza de rutina en su Banco de Sangre. Debe obtener una reacción de +++. 6.- Estos eritrocitos pueden ser guardados a 4ºC por 48 hrs.
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
MUESTRA: - Sangre sin anticoagulante, con no más de 48 hrs .de extraída. MATERIALES: -
Tubos de Khan (vidrio 12x 75 mm) Pipetas Pasteur con capuchón de goma. Gradillas para tubos. Centrífuga para inmunohematología previamente calibrada. Lector para Inmunohematología.
REACTIVOS: - Suero salino isotónico tamponado pH 7.0,o PBS. - Suero antiglobulina humana poliespecífico.
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
NOTA: a) si se dispone de un set de eritrocitos comercial, utilizar según las indicaciones del fabricante. b) El tiempo de incubación puede acortarse si en el punto 2 los eritrocitos se suspenden en medio de baja fuerza iónica, LISS, teniendo la precaución de utilizar los tiempos
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
incuba junto a los glóbulos rojos y plasma a probar para observar aglutinación. Luego de un proceso de centrifugación, las células que den reacción negativa (no aglutinación) pasaran por la matriz de gel hasta el fondo del tubo, en cambio las reacciones positivas (aglutinación) quedaran atrapadas en la matriz con diferentes intensidades según sea la reacción.
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
Doble población: Doble banda de hematíes, en el fondo y en la parte superior de la columna. Hemólisis: Sobrenadante y columna de color rosa.
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
INTERPRETACIÓN La presencia de anticuerpos se visualiza en forma de aglutinación o hemólisis en los tubos I y/o II. Para un resultado válido, el tubo + debe resultar positivo y el - debe ser negativo.
CONTROL DE CALIDAD:
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
7.- Lavar los tubos negativos o débilmente positivos por 4 veces con salino tamponado o PBS para realizar la prueba de AGH. 8.- Después del último lavado, eliminar totalmente el sobrenadante, agregar 1 gota de suero AGH poliespecífico y proceder como en los puntos 3 y 4. 9.- En los tubos que resultaron negativos, agregar 1 gota de células control AGH y proceder como en los puntos 3 y 4.
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
SANGRE Y COMPONENTES SANGUINEOS
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
PREPARACION Y CONSERVACION DE COMPONENTES SANGUINEOS SANGUINEOS Los anticoagulantes y soluciones preservadoras(ver tabla) que en la actualidad se añaden a la sangre del donante hacen posible su conservación por períodos más prolongados; esto junto al uso de sistemas cerrados de recolección que llevan adosados otras bolsas satélites de forma integral, permite la separación aséptica de los componentes sanguíneos. Las condiciones óptimas de velocidad, tiempo y temperatura requeridos tanto para la preparación como conservación y rendimiento de estos componentes, son diferentes para cada uno y deben estar determinadas para cada centrífuga y cada laboratorio en particular y revisarse periódicamente (ver tabla).
CONTENIDO DE LAS SOLUCIONES PRESERVADORAS (g/L) CONTENIDO Citrato trisódico Ácidocítrico Dextrosa Fosfato de Sodio adenina
ACD-A 22.00 8.00 24.50
CPD 26.30 3.27 25.50 2.22
CP2D 26.30 3.27 51.10 2.22
CPDA-1 26.30 3.27 31.90 2.22 0.275
CAMBIOS BIOQUIMICOS DE SANGRE CONSERVADA CON CPD Y CPDA-1 CPD VARIABLES SANGRE ST Dias conservación 0 21 0 % cel. viables a las 24 100 80 100 horas pH a 37ºC 7.2 6.84 7.6 % ATP (del valor 100 86 100 inicial) 2,3DPG (del valor 100 44 100 inicial) K+ plasma * 3.9 21.0 4.2 Na+ plasma * 168.0 156.0 169.0 Hb plasmática 17.0 191.0 82.0 * = mmol/L ; & =el plasma total de estas unidades es 70 ml.
CPDA-1 GR 0 100
ST 35 79
GR 35 71
7.55 100 100
6.98 56+/16 <10
6.71 45+/12 <10
5.1 169.0 78.0
27.3 155.0 461.0
78.5 & 111.0 658.0
PREPARACION DE COMPONENTES SANGUINEOS POR CENTRIFUGACIÓN. CENTRIFUGACIÓN. Centrifugación a "alta velocidad Concentrado de Glóbulos Rojos Concentrado de Plaquetas. Plasma Plasmaféresis Crioprecipitado
estandarizar cada centrifuga Ej.: 5000 x g, 7 minutos
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
METODOS A. CONCENTRADO DE GLOBULOS ROJOS. 1.
Extraer sangre en bolsa doble o triple y mantenerla en refrigeración a 4ºC hasta ser procesada. 2. Centrifugar a "alta velocidad" y a 5ºC. 3. Colocar la bolsa principal que contiene la sangre centrifugada en un compresor de plasma y soltar el muelle, dejando que la placa del compresor entre en contacto con la bolsa. 4. Obturar el tubo entre la bolsa primaria y las satélites con una pinza o hacer un nudo flojo. Si hay 2 bolsas satélites, colocar una pinza para que el plasma entre a sólo una de ellas. 5. Marcar las bolsa satélites con la misma identificación que la bolsa principal (Nº donante, fecha de preparación, tipo de componente ) y romper el sello. La eliminación de 225-250 ml de plasma dará lugar a unos hematíes residuales con 70- 80% de Hto. 6.- Volver a pinzar cuando haya entrado en la bolsa satélite la cantidad de plasma deseada. Sellar la tubuladura entre ambas bolsas en dos lugares y cortar en esa zona. 7.- Almacenar los componentes sanguíneos obtenidos a temperatura adecuada para cada uno. Si se extrae sangre en bolsa única: después de dejar la bolsa en el compresor, colocar una pinza en la tubuladura de la bolsa de trasferencia estéril, insertar en condiciones asépticas la cánula de la bolsa transfer en el orificio de salida de la bolsa de sangre, aflojar la pinza y continuar el procedimiento antes señalado. Esta unidad preparada en "circuito abierto" debe ser transfundida dentro de las 24 hrs.
B. CONCENTRADO DE GLOBULOS ROJOS POBRE EN LEUCOCITOS (bolsa invertida) 1.- Utilizar sangre con no más de 6 horas de extraída. 2.- Centrifugar la bolsa en forma invertida a "alta velocidad" y 5ºC. 3.- Colgar de un soporte la bolsa primaria que contiene la sangre centrifugada y continuar el procedimiento igual al anterior a partir del punto 4. Traspasar a la bolsa satélite o transfer estéril solo los dos tercios de los glóbulos rojos eliminado así cerca del 75 % de leucocitos. Si se extrae sangre en bolsa única: seguir las indicaciones arriba expuestas.
C. PLASMA FRESCO CONGELADO (PFC) 1.- Extraer sangre en bolsa doble o triple, mantenerla en refrigeración a 4ºC y procesarla antes de 6 hrs de extraída. 2.- Proceder igual a método A. o más, éste quede
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
D. CRIOPRECIPITADO DE GLOBULINA ANTI HEMOFILICA(AHF) O FACTOR VIII. 1.- Extraer sangre en bolsa doble o triple, mantenerla en refrigeración a 4ºC y procesarla antes de 6 hrs de extraída. 2.- Proceder igual a método A 3.- Congelar el plasma en forma rápida antes de 1 hr, esto se logra con congeladores de aire forzado o congeladores mecánicos capaces de mantener temperaturas de -65ºC o menos, hielo seco o un baño de etanol-hielo seco. 4.- Dejar descongelar el plasma a 1-6ºC colocando la bolsa en un refrigerador a 4ºC. 5.- Centrifugar el plasma a "alta velocidad" y 5ºC una vez descongelado. 6.- Colocar la bolsa en una prensa de plasma y dejar pasar a la bolsa satélite el 90% del plasma sobrenadante sin factor VIII dejando 10 a 15 ml de plasma para resuspender el crioprecipitado al momento de usar. 7.- Pinzar la tubuladura, comprobar que cada bolsa tenga la identificación correcta (Nº donante, fecha elaboración, tipo de componente). Sellar la tubuladura entre ambas bolsas en dos lugares y cortar en esa zona. 8.- Congelar rápidamente el crioprecipitado a -65ºC y almacenar a 20ºC. En estas condiciones dura 12 meses a partir de la fecha de extracción. Almacenar el plasma sin F VIII a - 20ºC o 4ºC
E. CONCENTRADO DE PLAQUETAS DE DONANTE AL AZAR: 1.- Extraer sangre en bolsa doble o triple, mantener a temperatura ambiente (22ºC) antes de separar el plasma rico en plaquetas de los hematíes. Procesar dentro de 6 hrs de extracción. 2.- Centrifugar mediante centrifugación "suave" a 20ºC para obtener plasma rico en plaquetas. 3.- Ídem método A, puntos 3 - 4 -5 4.- Traspasar el plasma rico en plaquetas a la bolsa satélite prevista como "Concentrado de Plaquetas" (CP). Evitar la contaminación con glóbulos rojos. Sellar la tubuladura en dos puntos entre la bolsa primaria y bolsa satélite CP y cortar entre ellos. Refrigerar los hematíes a 4ºC. 5.- Centrifugar el plasma rico en plaquetas a 20ºC y "alta velocidad". 6.- Traspasar el plasma sobrenadante pobre en plaquetas a otra bolsa satélite; dejar en el concentrado de plaquetas 50-60ml de plasma. Sellar el tubo en dos puntos ente ambas bolsa y cortar entre ellos. 7.- El concentrado plaquetario debe permanecer quieto a
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
CARACTERISTICAS DE LOS PRINCIPALES COMPONENTES SANGUINEOS COMPONENTE CONSERVACION VOLUMEN CONTENIDO (DIAS) (ML) GLOBULOS CPD: 21 250 GR: 65-80% ROJOS CPD-A: 35 a PLASMA: CONCENTRAHEPARINA: 2 300 20-35% DOS C.A.: 24 hr ALGUNAS: Tº:2-6ºC plaquetas y GB G.R. POBRES C.C.: igual 175 GR: 80-95% EN LEUCO GR a PLASMA: C.A.: 24 Hr 245 5-20% GB: < 30% Tº:2-6ºC PLAQUETAS Dadores al Azar
3-5 depende del plástico 24 Hrs (C.A.)
50 -65
PLAQUETAS: 5.5 x 1010 PLASMA: 50-65ml GB:
Tº:22ºC PLAQUETAS Donante Único
24 Hrs (C.A.) 3 -5 depende del plástico
300 a 500
PLASMA fresco congelado
1 año congelado 6-24 hr líquido
150 a 250
PLAQUETAS: 3.0 x 1011 PLASMA: 300-500 ml GB: * PLASMA: todos los F. de coagulación F.I: 400 mg
Tº: -20ºC PLASMA Conservado
CRIOPRECIPITADO
5 años congelado líquido: 5 díasmás que sangre Tº: -20ºC Congelado: 1 año Descongelad 6-8 hr En pool:
150 a 250
10-30
PLASMA: Solo F. estables
F.VIII C: 80-150 mg F.XIII: 20-30% del nivel st.
CONTROL DE CALIDAD - Hto: < 80% -Cultivo bacteriano según programa de Con.Cal. -Recuperación: 70-80% GR -Remoción: 70% GB (< 5x108 GB) -Hto:>o= 80% -Recuento: & 5.5x1010 Plaq. -pH: 6.0 o mas al fin de su conservación -Preparación: antes de 6 hrs. de extracción -Volumen: 50-65 ml -Recuento: & 3.0x1011 Plaq. -pH: 6.0 ídem -Preparación: antes de 6 hrs. de extracción -Conservación: < 18ºC - Descongelar: 30-37ºC -Conservación: < 18ºC
- 80 UI F.VIII en el 75% de las unidades estudiadas. (1 vez al mes)
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
C.A.: Circuito abierto, C.C.: Circuito cerrado GR:Glóbulos rojos, GB: Glóbulos blancos &: en el 75% de las unidades estudiadas.
CALIBRACION DE CENTRIFUGA PARA OBTENCION DE PLAQUETAS. 1.- Recoger un tubo de sangre del donante con EDTA. Realizar recuento de plaquetas. Si es inferior a 133 X 10 3/ ul no debe utilizar esta unidad para calibración. 2.- Calcular el Nº de plaquetas en la unidad de sangre total: recuento de plaq/ul X 1000 X ml de sangre = Nº de plaq en la unidad de sangre total. 3.- Centrifugar la unidad de sangre a velocidad y tiempo seleccionado ej.: 1200 rpm, 10 min. 4.- Obturar temporalmente la tubuladura de una bolsa satélite. Traspasar el plasma rico en plaquetas (PRP) según técnica. No retirar el obturador temporal hasta el paso siguiente. 5.- Exprimir la tubuladura varias veces hacia dentro de la bolsa para que contenga una muestra representativa del PRP. 6.- Sellar un segmento del tubo tubo y desconectarlo de manera que la bolsa de PRP siga siendo estéril. 7.- Realizar recuento plaquetario de esta muestra y calcular el Nº de plaquetas en la bolsa de PRP: Recuento plaquetas/ul x 1000 x ml de PRP = Nº de plaquetas en el PRP 8.- Calcular el porcentaje de recuperación: Nº de plaquetas en PRP X 100 = recuperación recupera ción (%) Nº plaquetas en sangre total 9.- Repetir el procedimiento anterior 3 o 4 veces con distintos donantes, utilizando centrífugas, velocidades y tiempos distintos. Comparar los rendimientos de cada ensayo 10.- Seleccionar el tiempo más corto y velocidad menor me nor que dé el mayor porcentaje de recuperación de plaquetas en PRP con un nivel aceptable de contaminación de rojos. 11.- Proceder a la preparación del concentrado plaquetario(CP) en la forma
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
15.- Repetir los pasos 11 a 14 con distintas centrífugas, velocidades y tiempos distintos. Comparar los rendimientos de cada conjunto de condiciones de centrifugación. 16.- Seleccionar el tiempo más corto y velocidad menor que den mayor porcentaje de recuperación de plaquetas en el concentrado. Una vez calibrada la centrífuga, no es necesario volver a hacerlo a no ser que se presente un problema de funcionamiento o que disminuya el rendimiento de los concentrados.
CONTENIDO APROXIMADO DE LEUCOCITOS EN LOS COMPONENTES SANGUINEOS COMPONENTE SANGUINEO Sangre total Concentrado de eritrocitos Eritrocitos lavados GR congelados y desglicerolizados Plaquetas por aféresis Concentrados plaquetarios
LEUCOCITOS POR UNIDAD 109 108 107 106 - 107 106 - 108 107
REDUCCION DE LEUCOCITOS EN LOS COMPONENTES SANGUINEOS: DEFINICIONES: Leucocitos/ul: Nº de células en un volumen de 1 ul, se determina por método de conteo microscópico directo. Leucocitos/ml: Para determinar leucocitos por ml, multiplique el Nº de leucocitos/ul por 1000. (1000ul = 1 ml) Contenido residual (efluente): Se define como en Nº total de leucocitos que permanecen en la unidad después de la filtración. También se llama contenido efluente y se puede calcular : Leucocitos/ul X 1000 X volumen de la unidad (ml). Log (logaritmo) de reducción Es la reducción de leucocitos que se logra por filtración, expresada en logaritmo. Log es el poder en que el número 10 debe aumentar en orden de producir un número dado. Ejemplo: log de 1000 = log 10 3 = 3; log de 10000 = log 104 = 4
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
5.000 (5 x 10 3) leucocitos/ul 3.-Recuento residual (efluente): 5 leucocitos/ul 4.-Cálculo: a.-Eficiencia % de reducción de leucocitos: (1 – 5/5000) * 100
= 99.9%
b.-Log de reducción: log 10leucocitos iniciales/ul leucocitos residuales/ul log 105000 = 3.00 5 c.-Contenido residual: leucocitos/ul * 1000 * Volumen de la unidad(ml) 5 * 1000 * 350 = 1.75 * 10 6 leuco/unidad
DATOS BIOLOGICOS DE ALGUNOS FACTORES DE COAGULACION FACTOR
FIBRINOGENO PROTROMBINA F-V F-X F - VII F - VIII F - IX F - XI F - XIII Von
CONCENTRA APROX. PLASMA (mg/ml) 2.8 0.13 0.068 0.012 0.0005 0.00024 0.005 0.006 0.029 0.006
VIDA MEDIA (Horas) 99+/-3 73+/-7 12-36 33 5 13+/-3 23+/-2 61 282 20-40
NIVEL RECOMEND PARA CIRUGIA 1mg/ml 40-50% 10-30% 10-40% 10-20% 30-100% 20-60% 20-30% 10% 20-50%
FRACCION INTRAVASCULAR
RECUPERACION
72% 58%
22%
83% 51% 100% 80-100% 24-54% 100% 50% 73%
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
UNIDAD EFECTOS ADVERSOS DE LA TRANSFUSION SANGUINEA
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
EFECTOS ADVERSOS DE LA TRANSFUSION SANGUINEA INTRODUCCION: La transfusión de sangre y sus componentes generalmente es una forma segura y eficaz de corregir déficit hematológico, sin embargo no está libre de riesgo debido a diferentes causas: ser un trasplante de un tejido liquido presencia de nuevas enfermedades transmisibles, incompatibilidades no pesquisable, donante en periodo de ventana de una enfermedad transmisible y no detectada por las actuales técnica de laboratorio, falla inherente a toda actividad humana etc. Los efectos adversos que pueden presentarse con la administración de sangre involucran aspecto y problemas más amplios que la común mente llamado " reacciones transfusionales". Alrededor de un 10% de los receptores transfundidos presentan reacciones; algunas son evitables pero otras no. El conocimiento de los efectos adversos a la transfusión es necesario y obligatorio para el Tecnólogo Médico que ejerce en un servicio de sangre, para su prevención, reconocimiento precoz, investigar las causas y adoptar las medidas correctoras correspondientes. correspondientes .
A.- REACCIONES TRANSFUNSIONALES TRANSFUNSION ALES INMEDIATAS Las reacciones transfusionales inmediatas generalmente ocurren dentro las dos primeras horas de la transfusión, son de varios tipos y pueden deberse a causas inmunológicas y no inmunológicas. Las causas inmunológicas incluyen incompatibilidad del receptor con los eritrocitos del donante, glóbulos blancos, plaquetas o componentes plasmáticos. En la mayoría de los casos, las reacciones agudas se deben a la presencia de anticuerpos del receptor dirigidos contra antígenos presentes en el producto transfundido. Menos frecuentemente puede ocurrir por anticuerpos incompatibles en el plasma del donante.
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
REACCION TRANSFUSIONAL HEMOLITICA Es causada por la reacción antígeno - anticuerpo que activa el complemento, los sistemas de coagulación y una respuesta endocrina, provocando un aumento de la destrucción de los eritrocitos. La hemólisis puede ser predominantemente intravascular, debida a anticuerpos IgM que fijan y activan la vía clásica del complemento hasta C9 provocando la ruptura de las células en el torrente sanguíneo y liberación de hemoglobina al plasma. Los anticuerpos IgG que no activan complemento (ej.: anti-Rh) o solo parcialmente hasta la etapa C3 (ej.: anti Kell), causan hemólisis extravascular, mediada por fagocitosis o citotoxicidad de los eritrocitos por las células del sistema retículo endotelial (SRE) en el hígado y bazo. Otras causas de hemólisis son el daño osmótico de eritrocitos por inyección accidental de agua en la circulación,contaminaciónbacteriana, sobrecalentamiento etc.
REACCION TRANSFUSIONAL HEMOLITICA AGUDA Es causada por anticuerpos IgM dirigidos contra antígenos eritrocitarios produciendo hemólisis intravascular; generalmente son el resultado de
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
dolor o malestar en el sitio de punción, enrojecimiento facial o leve alza de la temperatura. En pacientes que se encuentran anestesiados durante la transfusión, dos importantes signos revelan este tipo tipo de reacción: hipotensión a pesar de una adecuada reposición de fluidos, y sangramiento anormal. Los efectos clínicos varían varían grandemente. Por un lado, menos del 10% de los pacientes con esta incompatibilidad, desarrollan hemólisis intravascular aguda, seguida de falla renal y muerte; mientras en otros, los efectos clínicos no son fácilmente reconocibles. Es difícil establecer la frecuencia exacta con que ocurre este fenómeno; sin embargo es lejos la causa más común de reacción hemolítica aguda y de muerte inmediata después de la transfusión.
REACCION FEBRIL NO HEMOLÍTICA A.- Por anticuerpos HLA Es causada por anticuerpos HLA del receptor que reaccionan con el correspondiente antígeno ubicado en los glóbulos blancos del donante. El complejo antígeno - anticuerpo activa complemento y se une a monocitos activado liberando citoquinas con propiedades pirogénicas.
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
TRANSFUSION TRANSFUSION DE GLOBULOS ROJOS CONTAMINADOS Los glóbulos rojos contaminados con bacterias se reconocen por la presencia de hemólisis y la decoloración de ellos. Los efectos tóxicos inmediatos que produce esta sangre al ser transfundida provocan a veces un daño mayor que las otras reacciones hemolíticas. La contaminación de la sangre no es frecuente, pero ocurre por las siguientes razones: - Bacteremia asintomática en donantes aparentemente sanos con un pequeño número de bacterias, como por ej. Yersinia enterocolítica. - Contaminación de la unidad de sangre durante la recolección con bacterias de la flora normal de la piel del donante (ej.: staphylococcus epidermis, micrococcus o sarcina spp) - solución anticoagulante contaminada - Contaminación de los sellos de las bolsas de crioprecipitado o plasma congelado, al descongelarlos en agua infectada comúnmente con pseudomonas.
FALLA PULMONAR AGUDA (TRALI) Esta reacción es poco usual, pero potencialmente fatal. Ocurre inmediatamente después de la transfusión debido a anticuerpos leucocitarios del donante contra leucocitos del receptor o viceversa, produciéndose una leucoaglutinación .
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
HEMOLISIS CAUSADA POR UN EFECTO OSMOTICO Inyección accidental de agua en la circulación , conduce a una reacción hemolítica . Esto se presenta generalmente en operados de prostactectomia transuretral.
HEMOLISIS CAUSADA POR DAÑO FÍSICO DE LA SANGRE A.- Sobre calentamiento En algunas ocasiones es preciso entibiar la sangre para transfundirla, si la la temperatura excede los 37ºC puede producirse hemólisis de la unidad. Aparte de considerar las razones clínicas para entibiar la sangre, es necesario utilizar el equipo apropiado para esta operación, y verificar el control de temperatura de éste. En caso de utilizar un baño María, controlar la temperatura con un termómetro durante todo el proceso. B.- Congelación La unidad de sangre puede congelarse por almacenamiento en refrigerador no diseñado para esta función o dejada en el congelador en forma accidental desconocimiento de las
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
aumenta el título y avidez de éste se produce la hemólisis de los eritrocitos El segundo tipo tiene lugar en sujetos ya inmunizados, pero la concentración de anticuerpos es tan baja que no puede ser detectada por los tests pre transfusionales; o bien se produce una respuesta anamnéstica, elevándose la concentración de anticuerpos 3 días más tarde. Síntomas y signos más comunes de esta reacción Fiebre Ictericia leve Descenso de la Hemoglobina Hemoglobinuria
ENFERMEDAD INJERTO VERSUS HUESPED Es una complicación no habitual y tiene lugar en pacientes inmunodeprimidos, en los cuales los linfocitos inmunocompetentes del donante, anidan y se multiplican, reaccionando contra los tejidos “extraños” del huésped. El síndrome clínico puede incluir fiebre, erupción cutánea, diarrea,
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
CUADRO RESUMEN DE LOS EFECTOS ADVERSOS DE LA TRANSFUSIÓN SANGUÍNEA EFECTOS INMEDIATOS EFECTOS INMUNOLOGICOSETIOLOGIA HABITUAL Hemólisis sintomática
G. Rojo Incompatibles
Febril no Hemolítica
Granulocitos Donante
Anafiláctica
Ac Anti Ig A
Urticaria
Ac Anti Proteínas Plasma
Edema Pulmonar no cardiológico
Ac. Anti Leucocitos o Activación del C
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
EFECTOS RETARDADOS EFECTOS INMUNOLOGICOSETIOLOGIA HABITUAL Hemólisis sintomática
Ac. Anamnésicos eritrocitarios
Reacción injerto / huésped
Prolif. Linfocitos funcionales del donante
Púrpura post transfusional Aloinmunización contra ag. de eritrocitos Leucocitos, plaquetas, proteínas plasma
Anticuerpos anti plaquetarios
EFECTOS NO INMUNOLOGICOS
frente
antígenos
Exposición a antígenos del donante
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
INVESTIGACION DE UNA REACCION TRANSFUSIONAL
En toda UMT o Banco de Sangre o Servicio de Transfusiones, las reacciones transfusionales adversas se presentan de tiempo en tiempo, porque aunque el donante es compatible, no es posible tener todos los antígenos eritrocitarios, leucocitarios y plaquetarios, idénticos con el receptor. Las reacciones alérgicas y febriles son las más comunes y en muchos casos inevitables, al menos inicialmente. Las reacciones hemolíticas tardías ocurren con cierta frecuencia pero no siempre son fáciles de detectar o prevenir Las pruebas cruzadas solo sirven para reducir considerablemente la posibilidad de
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
Observar las condiciones de las unidades devueltas y filtro utilizado 1. - Efectuar investigación administrativa comprobando los datos del paciente y de la (s) unidad (es) con: -
los registros existentes en el banco de sangre muestra post transfusión muestra pre transfusión unidad (s)transfundida (s) informe de reacción adversa etiquetas de la unidad transfundida resultado de la prueba cruzada
Si hay discrepancia en este punto, en especial en la identificación nominal y/o de grupo sanguíneo, avisar inmediatamente al médico tratante y descartar que no estén otros pacientes involucrados, verificando las transfusiones hechas cercanas al
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
ENFERMEDADES DE TRANSMISION SANGUINEA
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
TESTS SEROLOGICOS EN EL DIAGNOSTICO DE HEPATITIS VIRAL AGENTE REACTIVIDAD DEL TEST HBsAg Anti-HBc AntiHBeAg Tot. IgM
INTERPRETACION INTERPRETACIO N Anti-
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
-
incubación a temperatura incorrecta Material sucio Mantener los reactivos en forma incorrecta Omisión de la muestra Registros incorrectos Error humano
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__________________________________________________________ _____________________________ ___________________________________________________INMUNOHEMATOLOGIA ______________________INMUNOHEMATOLOGIA Y MEDICINA TRANSFUSIONAL
•
La especificidad Es la capacidad de la prueba de dar un resultado negativo en personas que no presentan la enfermedad en el momento del estudio. Corresponde a la relación del número de verdaderos negativos sobre el total de sanos D/B+D.
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______________________________________________________ _________________________ _______________________________________________________INMUNOHEMATOLOGIA __________________________INMUNOHEMATOLOGIA Y MEDICINA MEDICINA TRANSFUSIONAL
BIBLIOGRAFIA 12.-
Race R. and Sanger "Blood groups in man"6th Ed. Neville J. Bryant "An introduction of immunohematology" 2th