Development of a varicella-zoster virus neutralization assay using a glycoprotein K antibody enzyme-linked immunosorbent spot assay.
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.
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian Province, China.
Lihong
L
Chen
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian Province, China.
Jian
J
Liu
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian Province, China.
Wei
W
Wang
National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, Xiamen 361102, Fujian Province, China.
Jianghui
J
Ye
Department of Obstetrics and Gynecology, the Affiliated Zhongshan Hospital of Xiamen University, Xiamen 361004, Fujian Province, China.
Lanling
L
Wen
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.
Qinjian
Q
Zhao
Department of Microbiology and Molecular Genetics, New Jersey Medical School, Rutgers University, 225 Warren Street, Newark, NJ 070101, USA.
Hua
H
Zhu
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.
Tong
T
Cheng
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.
Ningshao
N
Xia
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods; D006023:Glycoproteins / Q000276:immunology*; D014645:Herpesvirus 3, Human / Q000276:immunology*; D006801:Humans; D051379:Mice; D009500:Neutralization Tests / Q000379:methods*; D015203:Reproducibility of Results; D014759:Viral Envelope Proteins / Q000276:immunology*
D000914:Antibodies, Viral; D006023:Glycoproteins; C118859:ORF5 protein, Human herpesvirus 3; D014759:Viral Envelope Proteins
ELISPOT; Nutralization; Paque-reduction assay; VZV; gK