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Clinical and vaccine immunology : CVI2006; 14(2); 134-138; doi: 10.1128/CVI.00322-06

Monoclonal antibody-based competitive enzyme-linked immunosorbent assay for detecting and quantifying West Nile virus-neutralizing antibodies in horse sera.

Abstract: A rapid immunoassay for detecting and quantifying West Nile virus (WNV)-neutralizing antibodies in sera was developed as an alternative to the plaque reduction neutralization test (PRNT), the gold standard test for WNV. The assay is a competitive, enzyme-linked immunosorbent assay using neutralizing monoclonal antibody 5E8 (NT-ELISA). A cutoff percent inhibition (PI) value of 35% (mean PI plus 3 standard deviations), with a specificity of 99%, was established based on analysis of 246 serum samples from horses free of WNV. The NT-ELISA detected neutralizing antibodies in all sera collected 7 or 14 days postinoculation from mice (n = 11) infected with lineage I (strain NY385-99) or II (strain B956) WNV. When sera from WNV-vaccinated horses (n = 212) were tested by NT-ELISA and PRNT, the NT-ELISA gave a positive result for 96.1% (173/180) of the PRNT-positive sera and 3.1% (1/32) of the PRNT-negative sera. Discrepancies between the two tests were observed mainly with sera with low PRNT(90) titers (expressed as the reciprocal of the highest dilution yielding > or = 90% reduction in the number of plaques) for WNV or low PIs by NT-ELISA. The overall agreement (k value) between the two tests was 0.86. A good correlation (r(2) = 0.77) was also observed between the tests for endpoint titration of sera (n = 116). In conclusion, the newly developed NT-ELISA may be a good alternative serologic assay for detecting WNV that can be used for large-scale testing of WNV-neutralizing antibodies in multiple species.
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Publication Date: 2006-11-29 PubMed ID: 17135450PubMed Central: PMC1797797DOI: 10.1128/CVI.00322-06Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research is about a new immunoassay for detecting and quantifying West Nile virus (WNV) antibodies in horses serum. The assay, known as NT-ELISA, provides a quicker and more effective alternative to the plaque reduction neutralization test (PRNT) which is currently the gold standard for testing WNV.

Objective and Method

  • The research aimed to create a fast and efficient process, NT-ELISA, for detecting WNV antibodies in serum especially from horses. It was meant to offer a more effective alternative to PRNT.
  • The research involved developing a competitive enzyme-linked immunosorbent assay using the monoclonal antibody 5E8 for neutralizing WNV.

Test Validation

  • For validating this new assay, a cutoff percent inhibition value of 35% was set with a specificity of 99%, derived from the analysis of 246 horse serum samples free of WNV.
  • The NT-ELISA was tested on the sera taken from mice seven or 14 days after being inoculated with either lineage I or II WNV. It was able to detect neutralizing antibodies in all tested sera.

Comparison with PRNT

  • The researchers tested 212 horse serum samples vaccinated for WNV using both NT-ELISA and PRNT. NT-ELISA showed a positive result for 96.1% of PRNT positive sera and 3.1% of PRNT negative sera.
  • There were some discrepancies noticed between the results of two tests, especially with the sera which demonstrated low PRNT(90) titers or low PIs using NT-ELISA.
  • The overall agreement rate, denoted by k value, was relatively high at 0.86 indicating good agreement between the NT-ELISA and PRNT results.

Conclusion

  • The research concluded that NT-ELISA can be a reliable alternative for detecting and quantitating WNV-neutralizing antibodies. This assay could be particularly useful for large-scale testing involving multiple species.

Cite This Article

APA
Choi KS, Ko YJ, Nah JJ, Kim YJ, Kang SY, Yoon KJ, Joo YS. (2006). Monoclonal antibody-based competitive enzyme-linked immunosorbent assay for detecting and quantifying West Nile virus-neutralizing antibodies in horse sera. Clin Vaccine Immunol, 14(2), 134-138. https://doi.org/10.1128/CVI.00322-06

Publication

Clinical and vaccine immunology : CVI
ISSN: 1556-6811
NlmUniqueID: 101252125
Country: United States
Language: English
Volume: 14
Issue: 2
Pages: 134-138

Researcher Affiliations

Choi, Kang-Seuk
  • National Veterinary Research and Quarantine Service, 480 Anyang-6 dong, Anyang, Gyeonggi 430-824, Republic of Korea. choiks@nvrqs.go.kr
Ko, Young-Joon
    Nah, Jin-Ju
      Kim, Yong-Joo
        Kang, Shien-Young
          Yoon, Kyoung-Jin
            Joo, Yi-Seok

              MeSH Terms

              • Animals
              • Antibodies / blood
              • Antibodies, Monoclonal
              • Enzyme-Linked Immunosorbent Assay
              • Horses / blood
              • Horses / immunology
              • Serum / immunology
              • West Nile virus / immunology

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