Cargando…

Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum

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 ass...

Descripción completa

Detalles Bibliográficos
Autores principales: Jin, Jun, Ma, Hongxia, Xu, Lin, An, Dong, Sun, Shiyang, Huang, Xueyong, Kong, Wei, Jiang, Chunlai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7112850/
https://www.ncbi.nlm.nih.gov/pubmed/23178532
http://dx.doi.org/10.1016/j.jviromet.2012.11.014
_version_ 1783513556773240832
author Jin, Jun
Ma, Hongxia
Xu, Lin
An, Dong
Sun, Shiyang
Huang, Xueyong
Kong, Wei
Jiang, Chunlai
author_facet Jin, Jun
Ma, Hongxia
Xu, Lin
An, Dong
Sun, Shiyang
Huang, Xueyong
Kong, Wei
Jiang, Chunlai
author_sort Jin, Jun
collection PubMed
description 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 15 h 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.
format Online
Article
Text
id pubmed-7112850
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Elsevier B.V.
record_format MEDLINE/PubMed
spelling pubmed-71128502020-04-02 Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum Jin, Jun Ma, Hongxia Xu, Lin An, Dong Sun, Shiyang Huang, Xueyong Kong, Wei Jiang, Chunlai J Virol Methods Article 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 15 h 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. Elsevier B.V. 2013-02 2012-11-20 /pmc/articles/PMC7112850/ /pubmed/23178532 http://dx.doi.org/10.1016/j.jviromet.2012.11.014 Text en Copyright © 2012 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Jin, Jun
Ma, Hongxia
Xu, Lin
An, Dong
Sun, Shiyang
Huang, Xueyong
Kong, Wei
Jiang, Chunlai
Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum
title Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum
title_full Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum
title_fullStr Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum
title_full_unstemmed Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum
title_short Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum
title_sort development of a coxsackievirus a16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7112850/
https://www.ncbi.nlm.nih.gov/pubmed/23178532
http://dx.doi.org/10.1016/j.jviromet.2012.11.014
work_keys_str_mv AT jinjun developmentofacoxsackievirusa16neutralizationassaybasedonpseudovirusesformeasurementofneutralizingantibodytiterinhumanserum
AT mahongxia developmentofacoxsackievirusa16neutralizationassaybasedonpseudovirusesformeasurementofneutralizingantibodytiterinhumanserum
AT xulin developmentofacoxsackievirusa16neutralizationassaybasedonpseudovirusesformeasurementofneutralizingantibodytiterinhumanserum
AT andong developmentofacoxsackievirusa16neutralizationassaybasedonpseudovirusesformeasurementofneutralizingantibodytiterinhumanserum
AT sunshiyang developmentofacoxsackievirusa16neutralizationassaybasedonpseudovirusesformeasurementofneutralizingantibodytiterinhumanserum
AT huangxueyong developmentofacoxsackievirusa16neutralizationassaybasedonpseudovirusesformeasurementofneutralizingantibodytiterinhumanserum
AT kongwei developmentofacoxsackievirusa16neutralizationassaybasedonpseudovirusesformeasurementofneutralizingantibodytiterinhumanserum
AT jiangchunlai developmentofacoxsackievirusa16neutralizationassaybasedonpseudovirusesformeasurementofneutralizingantibodytiterinhumanserum