Antibodies to Bovine Serum Albumin in Human Sera: Problems and Solutions with Casein-Based ELISA in the Detection of Natural Japanese Encephalitis Virus Infections

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  • Konishi Eiji
    Department of International Health, Kobe University Graduate School of Health Sciences, Japan Center for Infectious Diseases, Kobe University Graduate School of Medicine, Japan
  • Kitai Yoko
    Department of International Health, Kobe University Graduate School of Health Sciences, Japan
  • Nishimura Koichi
    Division of Microbiology, Kumamoto Prefectural Institute of Public Health and Environmental Science, Japan
  • Harada Seiya
    Division of Microbiology, Kumamoto Prefectural Institute of Public Health and Environmental Science, Japan

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<p>An ELISA system for measuring antibodies to nonstructural protein 1 (NS1) of Japanese encephalitis virus has already been established. This system uses an ELISA diluent containing casein, instead of bovine serum albumin (BSA). During a survey, we found that 21 (21%) of 102 children aged 1–5 years, who had no history of Japanese encephalitis vaccination and were without detectable neutralizing antibodies, showed positive results with this ELISA system. Western blotting analysis showed that sera from 19 (91%) of these 21 subjects had antibodies to BSA, but not NS1. These sera reacted with BSA antigen remaining in immunoaffinity-purified NS1 antigen. One solution to this problem was to reduce the BSA level to ≤1% of the NS1 amount. Another was to use a control well sensitized with BSA with the same amount as that contained in the NS1 antigen preparation.<tt> </tt></p>

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