Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum.
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| Abstract |
Serum neutralizing antibody titers are indicative of protective immunity against Coxsackievirus A16 (CV-A16) and Enterovirus 71 (EV71), the two main etiological agents of hand, foot and mouth disease (HFMD), and provide the basis for evaluating vaccine efficacy. The current CV-A16 neutralization assay based on inhibition of cytopathic effects requires manual microscopic examination, which is time-consuming and labor-intensive. In this study, a high-throughput neutralization assay was developed by employing CV-A16 pseudoviruses expressing luciferase for detecting infectivity in rhabdomyosarcoma (RD) cells and measuring serum viral neutralizing antibodies. Without the need to use infectious CV-A16 strains, the neutralizing antibody titer against CV-A16 could be determined within 15h by measuring luciferase signals by this assay. The pseudovirus CV-A16 neutralization assay (pCNA) was validated by comparison with a conventional CV-A16 neutralization assay (cCNA) in testing 174 human serum samples collected from children (age <5 years). The neutralizing antibody titers determined by these two assays were well correlated (R(2)=0.7689). These results suggest that the pCNA can serve as a rapid and objective procedure for the measurement of neutralizing antibodies against CV-A16.
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| Authors | Jun Jin, Hongxia Ma, Lin Xu, Dong An, Shiyang Sun, Xueyong Huang, Wei Kong, Chunlai Jiang |
| Journal | Journal of virological methods (J Virol Methods) Vol. 187 Issue 2 Pg. 362-7 (Feb 2013) ISSN: 1879-0984 [Electronic] Netherlands |
| PMID | 23178532 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Validation Study) |
| Copyright | Copyright © 2012 Elsevier B.V. All rights reserved. |
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