Development of a high-throughput assay for measuring serum neutralizing antibody against enterovirus 71

Enterovirus 71 (EV71) is the main etiologic agent of hand, foot, and mouth disease (HFMD) and causes frequently severe neurological complications and mortality in young children. The serum neutralizing antibody response is the major indicator of EV71 infection and protective immunity. The current se...

Full description

Saved in:
Bibliographic Details
Published in:Journal of virological methods 2010-04, Vol.165 (1), p.42-45
Main Authors: Huang, Mei-Liang, Chiang, Pai-Shan, Luo, Shu-Ting, Liou, Guan-Yuan, Lee, Min-Shi
Format: Article
Language:English
Subjects:
Antibodies, Neutralizing - blood
Antibodies, Viral - blood
Antibody response
Assay development
Biological and medical sciences
Cell Line
Clinical Laboratory Techniques - methods
Cytopathogenic Effect, Viral
Enterovirus
Enterovirus 71
Enterovirus A, Human - growth & development
Enterovirus A, Human - immunology
Fundamental and applied biological sciences. Psychology
Humans
Mass Screening - methods
Microbiology
Neutralization Tests - methods
Neutralizing antibody
Sensitivity and Specificity
Seroepidemiology
Serum - immunology
Techniques used in virology
Virology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Enterovirus 71 (EV71) is the main etiologic agent of hand, foot, and mouth disease (HFMD) and causes frequently severe neurological complications and mortality in young children. The serum neutralizing antibody response is the major indicator of EV71 infection and protective immunity. The current serum neutralization test based on inhibition of cytopathic effect (Nt-CPE) requires manual microscopic examination, which is time-consuming and labor-intensive. In this study, a high-throughput neutralization assay which employs enzyme immunoassay for detecting growth of EV71 in Rhabdomyosarcoma (RD) cells and measuring serum neutralizing antibody (Nt-EIA) against EV71 was developed. RD cells infected with 100 TCID 50 of EV71 for 36–42 h had the best performance and were selected for Nt-EIA. One hundred and twenty human sera (59 negative sera, 61 positive sera) were measured for EV71 neutralization antibody titers by Nt-CPE and Nt-EIA. Neutralization antibody titers against EV71 determined by Nt-EIA had a high sensitivity (100%), specificity (94.9%) and agreement (97.5%) by a qualitative comparison with Nt-CPE. In the quantitative comparison, the correlation coefficient between Nt-EIA and Nt-CPE was 0.91 after log transformation. Overall, the Nt-EIA is a suitable alternative assay for the quantitation of EV71 neutralizing antibody to EV71.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2009.12.015