Quantitative Detection Of Rabies Virus To Establish

Rabies is one of the amphixenosis, it is a severe danger to human health. The case-fatality rate(CFR) is 100%. And from now on, there is no effective therapeutic tool, vaccine inoculation is more important to rabies. So the rabies vaccine efficiency is imperative. Now the experiment to determine the vaccine potency is NIH experiment, which is recommended by American National Institutes of Health (NIH). In this experiment, rabies vaccine is used to immune mice, according to the antibody potency in mice to estimate the vaccine potency. The test period is long, so it's application is restricted. In recent years, internal and abroad scientis establish many in vitro tests which to detect RV antigen, e.g. antibody binding test, SRD (single-radial-immunodiffusion), ELISA (enzyme immunoassay),and ELISA is the most well-rounded in these tests. Now ELISA is used by FAB when they evaluate the Chiron and Aventis rabies vaccine, in 2003 year, WHO indicate it is possible that the in vitro test to substitute the NIH test.This study establish in vitro test (ELISA) to detect RV antigen.The whole rabies virus particle is used to immune animals and to get polyclone antibody, the virius structure hasn't changed, its antigenicity and immunogenicity can be remained well, the aG rabies virus increasing in guinea-pig's brain is used to immune guinea-pig. So the non-specificity antibody can be decreased effectively, caprylic acid-ammonium sulfate salting-out purificate the antibody. This high-titer antibody is used to coat the EIA plate to establish in vitro test, which can capture the rabies virus antigen exactly and objectively. This test has fine specificity; it has no non-specificity reaction to other virus and groundsubstance in vaccine. After diluting rabies vaccine reference from National Institute For The Control Of Pharmaceutical And Biological Products (6.7IU/ml) , standard curve has been established. It's best linearity extent is 10-110 mIU/ml. r = 0.9913±0.0510 (95% confidence interval, n=20, S=0.135, Sx=0.030), sensibility is 5.3 mIU/ml. And the test has been validated and applied initially. This test cans detecte rabies virus generation in cell transfering in culture, and it can detect semi-product rabies vaccine immediately. According to national standard substance (6.7IU/ml) ,the test can detect rabies vaccines in market, the potency is 2.4—9.4IU/ml, excluding individual sample, these rabies vaccines potency can exceed 2.5 IU/dose.