Development of a varicella-zoster virus neutralization assay using a glycoprotein K antibody enzyme-linked immunosorbent spot assay

Lihong Chen; Jian Liu; Wei Wang; Jianghui Ye; Lanling Wen; Qinjian Zhao; Hua Zhu; Tong Cheng; Ningshao Xia

doi:10.1016/j.jviromet.2014.01.014

Development of a varicella-zoster virus neutralization assay using a glycoprotein K antibody enzyme-linked immunosorbent spot assay

J Virol Methods. 2014 May:200:10-4. doi: 10.1016/j.jviromet.2014.01.014. Epub 2014 Jan 31.

Authors

Lihong Chen¹, Jian Liu¹, Wei Wang¹, Jianghui Ye², Lanling Wen³, Qinjian Zhao⁴, Hua Zhu⁵, Tong Cheng⁶, Ningshao Xia⁴

Affiliations

¹ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian Province, China.
² National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, Xiamen 361102, Fujian Province, China.
³ Department of Obstetrics and Gynecology, the Affiliated Zhongshan Hospital of Xiamen University, Xiamen 361004, Fujian Province, China.
⁴ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian Province, China; National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, Xiamen 361102, Fujian Province, China.
⁵ Department of Microbiology and Molecular Genetics, New Jersey Medical School, Rutgers University, 225 Warren Street, Newark, NJ 070101, USA.
⁶ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian Province, China; National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, Xiamen 361102, Fujian Province, China. Electronic address: tcheng@xmu.edu.cn.

PMID: 24486923
DOI: 10.1016/j.jviromet.2014.01.014

Abstract

Plaque-reduction assays have been used to detect varicella-zoster virus (VZV)-neutralizing antibodies in sera for many decades. The current study characterized the mouse monoclonal antibody (MAb) 18A10, specific for VZV envelope glycoprotein K (gK), and applied this antibody to a new type of neutralization assay in the VZV field. The procedure is called the neutralization enzyme-linked immunosorbent spot (N-ELISPOT) assay and evolved from the VZV immunoperoxidase focus assay. Optimization of the assay involved defining the optimum combination of virus plaque-forming units (PFU) and antibody dilution, which were found to be 0-100 PFU and 1:200, respectively. Furthermore, the N-ELISPOT assay produced results consistent with that obtained for the plaque-reduction neutralization assay. Considering that the plaque-reduction neutralization assay is time-consuming and labor-intensive, the VZV N-ELISPOT assay offers several advantages including reproducibility and applicability for high-throughput analysis of humoral immune responses to VZV.

Keywords: ELISPOT; Nutralization; Paque-reduction assay; VZV; gK.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Viral / blood*
Enzyme-Linked Immunospot Assay / methods
Glycoproteins / immunology*
Herpesvirus 3, Human / immunology*
Humans
Mice
Neutralization Tests / methods*
Reproducibility of Results
Viral Envelope Proteins / immunology*

Substances

Antibodies, Viral
Glycoproteins
ORF5 protein, Human herpesvirus 3
Viral Envelope Proteins