» Articles » PMID: 26831405

Protein Microarray-mediated Detection of Antienterovirus Antibodies in Serum

Overview
Journal J Int Med Res
Publisher Sage Publications
Specialty General Medicine
Date 2016 Feb 3
PMID 26831405
Authors
Affiliations
Soon will be listed here.
Abstract

Objective: To utilize prokaryotic gene expression and protein microarray to develop and evaluate a sensitive, accurate protein microarray assay for detecting antienterovirus antibodies in serum samples from patients with hand, foot and mouth disease (HFMD). Enterovirus 71 (EV71) and coxsackievirus A16 (CA16), two common causative agents for HFMD, were used for assay development.

Methods: Serum was collected from patients with HFMD and healthy controls. EV71 and CA16 VP1 and VP3 genes were expressed in transfected Escherichia coli; the resultant VP1 and 3 proteins were used in a microarray assay for human serum EV71 and CA16 immunoglobulin (Ig) M and IgG. To validate the microarray assay, serum samples were tested for EV71 IgM using enzyme-linked immunosorbent assay (ELISA).

Results: Out of 50 patients with HFMD, EV71 IgM and CA16 IgM was detected in 80% and 44% of serum samples, respectively, using protein microarray, and EV71 IgM was detected in 78% of samples using ELISA. Protein microarray and ELISA showed 100% specificity for EV71-IgM detection.

Conclusion: The protein microarray assay developed in the present study shows potential as a sensitive technique for detecting EV71 IgM in serum samples from patients with HFMD.

References
1.
Jiang W, Huang R, Duan C, Fu L, Xi Y, Yang Y . Identification of five serum protein markers for detection of ovarian cancer by antibody arrays. PLoS One. 2013; 8(10):e76795. PMC: 3792870. DOI: 10.1371/journal.pone.0076795. View

2.
Wu T, Wang Y, Lee Y, Wang J, Liu C, Wang S . Immunity to avirulent enterovirus 71 and coxsackie A16 virus protects against enterovirus 71 infection in mice. J Virol. 2007; 81(19):10310-5. PMC: 2045469. DOI: 10.1128/JVI.00372-07. View

3.
Gnjatic S, Wheeler C, Ebner M, Ritter E, Murray A, Altorki N . Seromic analysis of antibody responses in non-small cell lung cancer patients and healthy donors using conformational protein arrays. J Immunol Methods. 2008; 341(1-2):50-8. DOI: 10.1016/j.jim.2008.10.016. View

4.
Cui S, Wang J, Fan T, Qin B, Guo L, Lei X . Crystal structure of human enterovirus 71 3C protease. J Mol Biol. 2011; 408(3):449-61. PMC: 7094522. DOI: 10.1016/j.jmb.2011.03.007. View

5.
Wu J, Zhao N, Pan H, Wang C, Wu B, Zhang H . Patterns of polymorphism and divergence in the VP1 gene of enterovirus 71 circulating in the Asia-Pacific region between 1994 and 2013. J Virol Methods. 2013; 193(2):713-28. DOI: 10.1016/j.jviromet.2013.07.051. View