title: Development and evaluation of a pseudovirus-luciferase assay for rapid and quantitative detection of neutralizing antibodies against enterovirus 71.
abstract: The level of neutralizing antibodies (NtAb) induced by vaccine inoculation is an important endpoint to evaluate the efficacy of EV71 vaccine. In order to evaluate the efficacy of EV71 vaccine, here, we reported the development of a novel pseudovirus system expression firefly luciferase (PVLA) for the quantitative measurement of NtAb. We first evaluated and validated the sensitivity and specificity of the PVLA method. A total of 326 serum samples from an epidemiological survey and 144 serum specimens from 3 clinical trials of EV71 vaccines were used, and the level of each specimen's neutralizing antibodies (NtAb) was measured in parallel using both the conventional CPE-based and PVLA-based assay. Against the standard neutralization assay based on the inhibition of the cytopathic effect (CPE), the sensitivity and specificity of the PVLA method are 98% and 96%, respectively. Then, we tested the potential interference of NtAb against hepatitis A virus, Polio-I, Polio-II, and Polio-III standard antisera (WHO) and goat anti-G10/CA16 serum, the PVLA based assay showed no cross-reactivity with NtAb against other specific sera. Importantly, unlike CPE based method, no live replication-competent EV71 is used during the measurement. Taken together, PVLA is a rapid and specific assay with higher sensitivity and accuracy. It could serve as a valuable tool in assessing the efficacy of EV71 vaccines in clinical trials and disease surveillance in epidemiology studies.
authors.affiliation: Department of Microbiology and Infectious Disease Center, Peking University Health Science Center, Beijing, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Wu
authors.forename: Qunying
authors.initials: Q
authors.lastname: Mao
authors.forename: Xin
authors.initials: X
authors.lastname: Yao
authors.forename: Pan
authors.initials: P
authors.lastname: Chen
authors.forename: Xiangmei
authors.initials: X
authors.lastname: Chen
authors.forename: Jie
authors.initials: J
authors.lastname: Shao
authors.forename: Fan
authors.initials: F
authors.lastname: Gao
authors.forename: Xiang
authors.initials: X
authors.lastname: Yu
authors.forename: Fengcai
authors.initials: F
authors.lastname: Zhu
authors.forename: Rongcheng
authors.initials: R
authors.lastname: Li
authors.forename: Wenhui
authors.initials: W
authors.lastname: Li
authors.forename: Zhenglun
authors.initials: Z
authors.lastname: Liang
authors.forename: Junzhi
authors.initials: J
authors.lastname: Wang
authors.forename: Fengmin
authors.initials: F
authors.lastname: Lu
mesh_terms: D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood*; D045744:Cell Line, Tumor; D002675:Child, Preschool; D002986:Clinical Trials as Topic; D029821:Enterovirus A, Human / Q000276:immunology* / Q000502:physiology; D005260:Female; D006232:Hand, Foot and Mouth Disease / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D006801:Humans; D007223:Infant; D049409:Luciferases, Firefly / Q000096:biosynthesis; D008297:Male; D009500:Neutralization Tests; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014611:Vaccination; D014765:Viral Vaccines / Q000276:immunology; D053586:Virus Internalization
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D014765:Viral Vaccines; D049409:Luciferases, Firefly