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1993, Número 3

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Vet Mex 1993; 24 (3)


Obtención de anticuerpos contra progesterona y estradiol, estandarización del radioinmunoanálisis y validación en suero de rumiantes

Herrera DMR, Luna MM, Romero RCM

Idioma: Español/Inglés
Referencias bibliográficas: 37
Paginas: 223-230
Archivo PDF: 164.01 Kb.


RESUMEN

Para obtener anticuerpos específicos contra progesterona (P4) y 17ß-estradiol (E2) y la estandarización de sus radioinmunoensayos (RIAs) correspondientes, se inmunizaron conejos Nueva Zelanda y California con 100 o 160 µg de Progesterona-CMO:BSA o 17ß-EstradiolCMO:BSA respectivamente.Las reinoculaciones se realizaron con intervalo de una semana en seis ocasiones, titulándose en cada una el suero cosechado. Con base en el mejor título, se seleccionaron los antisueros, a los que se les determinó la especificidad mediante reacciones cruzadas con varios esteroides. La reacción cruzada de los antisueros obtenidos contra P4 fue de 4.4 y 4.4% para pregnenolona, de 4.0 y 13.3% para epipregnenolona y menos del 1.0% para alopregnenolona, testosterona, E2 y cortisol para AcQKP5 y AcQKP6 respectivamente. La reacción cruzada del antisuero obtenido contra E2 fue de 3.6% para estrona y menos de 0.01 % para estriol, androstendiona, 17α-estradiol, P4 y testosterona. La sensibilidad de los ensayos permite detectar desde 6.25 pg/tubo de P4 y 0.5 pg/tubo de E2. Los coeficientes de variación intra e interensayo en el RIA fueron para P4 de 10.9% y 12.8% y para E2 de 5.0 y 10.0% respectivamente. Para validar los RIAs se determinó la concentración de P4 y E2 en cabras y vacas en diferentes estados reproductivos. Así, la concentración de P4 en cabras durante el último tercio de gestación fue de 8.3 ± 0.4 ng/ml y durante el periodo preparto disminuyó a 5.4 ± 0.8 ng/ml; en esta última etapa el E2 mostró valores de 55.1 ± 8.1 pg/ml. En la vaca, los valores de P4 y E2 fueron en el estro de 0.9±0.08 ng/ml y 10.6 ± 2.4 pg/ml, en la fase lútea de 8.6 ± 2.7 ng/ml y 3.3 ± 0.6 pg/ml y en la gestación de 6.4 ± 0.6 ng/ml y 17.3 ± 1.6 pg/ml respectivamente. La sensibilidad de los RIAs con los anticuerpos obtenidos, es capaz de identificar y diferenciar los valores característicos de P4 y E2 de los diferentes estados reproductivos en rumiantes domésticos.


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