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Development and Validation of a Competitive ELISA Based on Bovine Monoclonal Antibodies for the Detection of Neutralizing Antibodies against Foot-and-Mouth Disease Virus Serotype A.

Yimei CaoKun LiXiangchuan XingGuoqiang ZhuYuanfang FuHuifang BaoXingwen BaiPu SunPinghua LiJing ZhangXueqing MaJian WangZhixun ZhaoDong LiZaixin LiuZengjun Lu
Published in: Journal of clinical microbiology (2022)
The level of neutralizing antibodies in vaccinated animals is directly related to their level of protection against a virus challenge. The virus neutralization test (VNT) is a "gold standard" method for detecting neutralizing antibodies against foot-and-mouth disease virus (FMDV). However, VNT requires high-containment facilities that can handle live viruses and is not suitable for large-scale serological surveillance. In this study, a bovine broadly neutralizing monoclonal antibody (W145) against FMDV serotype A was successfully produced using fluorescence-based single-B-cell antibody technology. Using biotinylated W145 as a detector antibody and another bovine cross-reactive monoclonal antibody, E32, which was produced previously as a capture antibody, a competitive enzyme-linked immunosorbent assay for the detection of neutralizing antibodies (NAC-ELISA) against FMDV serotype A was developed. The specificity and sensitivity of the assay were evaluated to be 99.04% and 100%, respectively. A statistically significant correlation ( r = 0.9334, P < 0.0001) was observed between the NAC-ELISA titers and the VNT titers, suggesting that the NAC-ELISA could detect neutralizing antibodies against FMDV serotype A and could be used to evaluate protective immunity.
Keyphrases
  • dengue virus
  • monoclonal antibody
  • disease virus
  • zika virus
  • aedes aegypti
  • transcription factor
  • high throughput
  • public health
  • escherichia coli
  • label free
  • drug induced
  • multidrug resistant
  • structural basis