Determination of critical factors causing cytotoxicity in the virus neutralization test

•The critical steps of the FAVN test that cause the cytotoxicity were determined.•The factors that cause the cytotoxicity in the FAVN test were determined.•The effective ways that prevent the cytotoxicity in the FAVN test were established.•The FAVN test could be modified to increase accuracy and per...

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Bibliographic Details
Published in:Journal of virological methods 2014-04, Vol.199, p.46-52
Main Authors: Shiraishi, Rikiya, Nishimura, Masaaki, Nakashima, Ryuji, Enta, Chiho, Hirayama, Norio
Format: Article
Language:English
Subjects:
Animals
Antibodies, Neutralizing - blood
Antibodies, Viral - blood
Cell Line
Cricetinae
Cytotoxicity
Dog Diseases - diagnosis
Dog serum
Dogs
False Negative Reactions
Female
Humans
Male
Neutralization Tests - methods
Rabies
Rabies - diagnosis
Rabies - veterinary
Rabies virus - immunology
Virus neutralization test
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Summary:•The critical steps of the FAVN test that cause the cytotoxicity were determined.•The factors that cause the cytotoxicity in the FAVN test were determined.•The effective ways that prevent the cytotoxicity in the FAVN test were established.•The FAVN test could be modified to increase accuracy and performance. The fluorescent antibody virus neutralization (FAVN) test, an international standard method for serological testing for rabies, has been adopted by many countries. However, some dog serum samples inhibit the formation of cell monolayers by BHK-21 cells used in the test, resulting in failures to determine antibody titers. This inhibition of cell monolayer formation was defined as cytotoxicity. In this study, critical factors that induce cytotoxicity of the dog serum in BHK-21 cells were determined, and the effective ways to prevent cytotoxicity were also established. Specifically, some anticoagulants, anti-BHK-21 cell IgG antibodies, and serum storage at temperatures of >25°C were found to induce cytotoxicity. On the other hand, several treatments of the dog serum, including the absorption by BHK-21 cells or kaolin, incubation with trypsin-EDTA, and the use of collagen- or gelatin-coated plates, were shown to reduce cytotoxicity. Based on these results, the FAVN test may be modified to enhance its performance.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2014.01.006