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Revista do Instituto de Medicina Tropical de São Paulo

Print version ISSN 0036-4665

Rev. Inst. Med. trop. S. Paulo vol.35 no.6 São Paulo Nov./Dec. 1993

http://dx.doi.org/10.1590/S0036-46651993000600011 

IMUNOLOGIA

 

Evaluacion de la tecnica de contraimmunoelectroforesis para determinar la potencia antigena de las vacunas antirrabicas

 

Evaluation ot the counterimmunoelectrophoresis technique to determine the antigenic potency of antirrabic vaccines

 

 

Graciela MiceliI; Jorge E. TorrobaII; Ana María DiazII

ICátedra de Inmunología Veterinaria. Facultad de Ciencias Veterinarias. UNLP. 60 y 118 - (1900) La Plata - Buenos Aires - Argentina
IIInstituto Panamericano para la Protección de Alimentos y Zoonosis (PAHO/WHO) - Talcahuano 1660 - (1640) Martínez - Buenos Aires - Argentina

 

 


RESUMEN

El método recomendado por la Organización Mundial de la Salud (OMS) para la prueba de potencia de vacunas antirrábicas como producto final es la prueba NIH. Algunas técnicas in vitro se han propuesto para el control durante el proceso de produción y complementan el ensayo in vivo antes mencionado.
Este trabajo presenta los resultados obtenidos cuando se utilizó la técnica de contrainmunoelectroforésis (CIE) para determinar el contenido de antígenos en muestras de 84 y 40 lotes de vacunas antirrábicas producidas en tejido nervioso de cerebro de ratón lactante mediante cultivo de tejidos, respectivamente.
La evaluación de las muestras en, y en torno de, las 0.3 UI por ambos métodos muestran que, en la práctica, un título CIE de 1:4 cumpliría con un mínimo de potencia de la prueba NIH. Un bajo grado de variabilidad de la prueba CIE fue observada en nuestro laboratorio cuando dos lotes de vacunas de referencia de trabajo y 7 lotes de vacunas antirrábicas, de diferente origen y actividad, fueron ensayadas en cinco pruebas independientes. Todos los títulos se ubicaron dentro de una dilución doble, lo que es indicativo de su reproducibilidad.
Se observó buena sensibilidad para detectar el deterioro del antígeno en el ensayo de degradación térmica, cuando muestras de 3 lotes de vacuna líquida de cerebro de ratón lactante fueron mantenidas a4 y 37ºC cada una, por 28 días. Se evaluaron semanalmente por los ensayos de CIE y NIH. Finalmente, se observó que el ensayo de CIE podría ser utilizado por los productores para estimar el punto final de los procesos de concentración para que se corresponda con un valor antigénico deseado en la prueba de potencia NIH.

Unitermos: Rabia; Vacunas; Control de potencia "in vitro"


SUMMARY

The method recommended by the World Health Organization (WHO) for the potency assay of human and animal rabies vaccines as final product is the NIH test. Some in vitro techniques have been proposed for in process testing and supplement the in vivo test mentioned above.
This paper presents the results obtained when using a counterimmunoelectrophoresis (CIE) techniques to determine the antigens content in samples of 84 and 40 samples of lots of suckling mouse rabies and tissue culture rabies vaccines, respectively.
The evaluation of rabies vaccines under, at, near and up to 0.3 UI by the NIH and CIE tests showed that for practical use, a CIE titer of 1:4 would match the minimum of NIH potency test requirement. Low degree of CIE test variability within our laboratory was observed, when two working reference and 7 lots of rabies vaccines from different origin and activity were tested in five independent assays. All titres fell within one double dilution what is an indication of its reproducibility.
Good sensitivity to detect antigen deterioration in thermal degradation test was observed when samples of 3 lots of liquid suckling mouse rabies vaccine were held stored at 4ºC or at 37ºC for 4 weeks and assays by CIE and NIH tests. Finally, CIE test could apply by manufactures to follow the concentration procedures to determine the final end point which could match a desired antigenic value in the NIH potency test.


 

 

Texto completo disponible sólo en PDF.

Full text available only in PDF format.

 

 

AGRADECIMIENTOS

Los autores agradecen a la Dra. Isabel N. de Kantor sus valiosas sugerencias en la preparación de este manuscrito.

 

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Recebido para publicação em 01/12/1992
Aceito para publicação em 22/07/1993