title: Development of a coxsackievirus A16 neutralization test based on the enzyme-linked immunospot assay.
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.
authors.affiliation: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, 361102, China.
authors.forename: Wangheng
authors.initials: W
authors.lastname: Hou
authors.affiliation: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, 361102, China.
authors.forename: Lisheng
authors.initials: L
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, 361102, China.
authors.forename: Delei
authors.initials: D
authors.lastname: He
authors.affiliation: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, 361102, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zheng
authors.affiliation: National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Longfa
authors.initials: L
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, 361102, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, 361102, China.
authors.forename: Yajing
authors.initials: Y
authors.lastname: Liu
authors.affiliation: National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Beijing Wantai Biological Pharmacy Enterprise Co., Ltd., Beijing, 102206, China.
authors.forename: Xiangzhong
authors.initials: X
authors.lastname: Ye
authors.affiliation: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, 361102, China; National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China. Electronic address: tcheng@xmu.edu.cn.
authors.forename: Tong
authors.initials: T
authors.lastname: Cheng
authors.affiliation: State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen, 361102, China; National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Ningshao
authors.initials: N
authors.lastname: Xia
mesh_terms: D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood*; D004770:Enterovirus / Q000276:immunology*; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods*; D057166:High-Throughput Screening Assays; D006801:Humans; D009500:Neutralization Tests / Q000379:methods*; D013997:Time Factors
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral
keywords: Antibody; Coxsackievirus A16; ELISPOT; Enterovirus; Neutralization