Estudo comparativo entre testes de controle de qualidade de vacinas anti-rabicas inativadas para uso animal e produção experimental da vacina de referencia nacional

by Martins Arita, Gonçala Maria

Abstract (Summary)
The efficiency of anti-rabies inactivated vaccines that are commercialized for animal use, in Brazil, is evaluated by the Habel test, considering ?1 04,o/DL 50 for official approval of the lots. The goals of the present study is to compare the NIH ("National Institute of Health") and the indirect ELISA tests to the Habel test and produce the National Reference Vaccine (VRN) experimentally, to be used in the NIH. Two hundred and ten 10ts of vaccine have been tested by the NIH test parallel to the Habel testo The NIH test, following the limits 0,9UI; 0,97UI; 0,98UI; 1,OUI and 1,1 UI approved, respectively, 77,62%; 75,71%; 75,23%; 73,81%; 71,90%, while the Habel test approved 82,38% ofthe 10ts. The cut offvalue determined for the NIH was more than 0,98 DI so 1,0 UI is recomended as the official cut-offvalue. The indirect ELISA was applied in seras of mice immunized with 70 lots of vaccines with varied results in the Habel. The cut off values, the sensibility (8), the specificity (E), the predictable values for positive (VPRP) and negative (VPRN) results and the ratio between positive standard sera and no vaccinated mice sera were determined, so it was decided to adopt in this method the dilution of 1/400 that presents cut-off value of 0,225; 8=97%; E=1O0%; VPRP=100% and VPRN=78%. The National Reference Vaccine (VRN) was produced in BHK cells, C 13, with the Pasteur vírus (PV). The viral suspensions (5!!! BHK) had been titres of 107,66DL 50/rnL in lactents mice and 107,17 DL 50/rnL in young adult mice and 3,10 _g/rnL of total rabies glycoproteins (G). The final volume was divided into samples A and B, tOOt were inactivated, respectively, by binary ethyleneimine (BEI) and by p- propiolactone (BPL). For both inactivants there was a fast decrease ofthe infectivity and a slow decrease ofviral degradation in G contento Sample A (BEI-12 hours) and B (BPL-6 hours) presented insoluble G=1,92 µg/rnL and 2,02 µg/rnL, respectively, as the contents. Both presented satisfactory results in the viral inactivation, in toxicity and in sterility tests. For the potency test, samples A and B were sub-divided into AI and BI (5x concentrated) and into A2 and B2 (l0x concentrated), that presented good response in quality control tests and were capable of inducing immune response in mice, mesured through IgG quantification and protection against the viral challenge, suggesting that the production methodology was adequate to the VRN obtention, compatible to the Regional Reference Vaccine -INPPAZ-, able to be amplified to supply the national request, in the NIH
This document abstract is also available in Portuguese.
Bibliographical Information:

Advisor:Clarice Weis Arns; Clarice Weis Arns [Orientador]; Sergio Marangoni; Cleide Aschenbrenner Consales; Aramis Augusto Pinto; Fernanda Ramos Gadelha

School:Universidade Estadual de Campinas

School Location:Brazil

Source Type:Master's Thesis

Keywords:vacinas controle de qualidade teste imunoenzimatico


Date of Publication:12/13/2002

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