title: A rapid and quantitative assay for measuring neutralizing antibodies of Coxsackievirus B3.
abstract: Coxsackievirus B3 (CVB3) infection has been found to account for an increasing proportion cases of hand, foot and mouth disease (HFMD) in recent epidemiology studies. CVB3 is a single stranded, non-enveloped RNA virus and the infection can cause prominent health threat to pre-school children. Here, by taking approaches of reverse genetics, we established a single-round infection system for CVB3. The pseudovirus was produced by sequential transfection of CVB3 capsid expresser plasmid and CVB3 replicon RNA bearing firefly luciferase as a reporter. The CVB3 pseudovirus system was used for quantifying neutralizing antibody (NtAb) levels of 720 human serum samples and showed superior specificity and sensitivity comparing traditional cytopathic effect (CPE) assay. Furthermore, we compared the seroprevalence of CVB3 NtAbs in pre-school children and healthy adults, and found that only 11.94% of pre-school children were NtAbs positive which suggested that most children were naive to CVB3 infection; while there is much higher positive rate in adults (60%) indicating that most adults have experienced CVB3 infection during childhood. This rapid and quantitative assay greatly facilitates evaluating the level of NtAbs against CVB3 in populations and will help to advance CVB3 vaccine development.
authors.affiliation: National Institutes for Food and Drug Control, Beijing 100050, China; National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Pan
authors.initials: P
authors.lastname: Chen
authors.affiliation: National Institutes for Food and Drug Control, Beijing 100050, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Wu
authors.affiliation: National Institutes for Food and Drug Control, Beijing 100050, China.
authors.forename: Qunying
authors.initials: Q
authors.lastname: Mao
authors.affiliation: National Institutes for Food and Drug Control, Beijing 100050, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Gao
authors.affiliation: National Institutes for Food and Drug Control, Beijing 100050, China.
authors.forename: Xiaotian
authors.initials: X
authors.lastname: Hao
authors.affiliation: National Institutes for Food and Drug Control, Beijing 100050, China.
authors.forename: Lianlian
authors.initials: L
authors.lastname: Bian
authors.affiliation: Jiangsu Provincial Center for Disease Prevention and Control, Nanjing 210009, China.
authors.forename: Fengcai
authors.initials: F
authors.lastname: Zhu
authors.affiliation: National Institute of Biological Sciences, Beijing 102206, China.
authors.forename: Wenhui
authors.initials: W
authors.lastname: Li
authors.affiliation: National Institutes for Food and Drug Control, Beijing 100050, China.
authors.forename: Miao
authors.initials: M
authors.lastname: Xu
authors.affiliation: National Institutes for Food and Drug Control, Beijing 100050, China. Electronic address: lzhenglun@126.com.
authors.forename: Zhenglun
authors.initials: Z
authors.lastname: Liang
mesh_terms: D000328:Adult; D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood*; D029822:Enterovirus B, Human / Q000235:genetics / Q000276:immunology*; D006232:Hand, Foot and Mouth Disease / Q000175:diagnosis* / Q000453:epidemiology; D006801:Humans; D007223:Infant; D008875:Middle Aged; D059386:Reverse Genetics; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies; D013997:Time Factors; D014773:Virology / Q000379:methods; D055815:Young Adult
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral
keywords: Coxsackievirus B3; Neutralizing antibody; Pseudovirus; Seroprevalence