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Development of a coxsackievirus A16 neutralization test based on the enzyme-linked immunospot assay.

Development of a coxsackievirus A16 neutralization test based on the enzyme-linked immunospot assay.

Authors :
Hou W
Yang L
He D
Zheng J
Xu L
Liu J
Liu Y
Zhao H
Ye X
Cheng T
Xia N
Source :
Journal of virological methods [J Virol Methods] 2015 Apr; Vol. 215-216, pp. 56-60. Date of Electronic Publication: 2015 Feb 19.
Publication Year :
2015

Abstract

Coxsackievirus A16 (CA16) is one of the major pathogens responsible for hand, foot and mouth disease (HFMD). The assessment of the humoral immunity response is indispensable in the development of vaccines against enteroviruses. The neutralization test based on the inhibition of cytopathic effects (Nt-CPE) is a common method for measuring neutralizing antibodies against CA16. However, an efficient neutralization test needs to be developed for seroepidemiological surveys and clinical trials of CA16 vaccines because Nt-CPE is time-consuming and labor-intensive. In this study, a high-throughput neutralization test for CA16 based on the enzyme-linked immunospot assay (Nt-ELISPOT) was developed. The monoclonal antibody 7D10, which reacted with the viral protein VP1, was used to detect the cells infected with CA16. The neutralizing titers of sera were proven to be unchanged over an infectious dose range from 10 to 10,000TCID50 per well. The Nt-ELISPOT results correlated well with the Nt-CPE results (R(2) = 0.9250), and the detection period was shortened from five days to approximately 30h. Overall, the Nt-ELISPOT is a reliable and efficient method for measuring neutralizing antibodies against CA16.<br /> (Copyright © 2015 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1879-0984
Volume :
215-216
Database :
MEDLINE
Journal :
Journal of virological methods
Publication Type :
Academic Journal
Accession number :
25704596
Full Text :
https://doi.org/10.1016/j.jviromet.2015.02.010