title: Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum.
abstract: Serum neutralizing antibody titers are indicative of protective immunity against Coxsackievirus A16 (CV-A16) and Enterovirus 71 (EV71), the two main etiological agents of hand, foot and mouth disease (HFMD), and provide the basis for evaluating vaccine efficacy. The current CV-A16 neutralization assay based on inhibition of cytopathic effects requires manual microscopic examination, which is time-consuming and labor-intensive. In this study, a high-throughput neutralization assay was developed by employing CV-A16 pseudoviruses expressing luciferase for detecting infectivity in rhabdomyosarcoma (RD) cells and measuring serum viral neutralizing antibodies. Without the need to use infectious CV-A16 strains, the neutralizing antibody titer against CV-A16 could be determined within 15h by measuring luciferase signals by this assay. The pseudovirus CV-A16 neutralization assay (pCNA) was validated by comparison with a conventional CV-A16 neutralization assay (cCNA) in testing 174 human serum samples collected from children (age <5 years). The neutralizing antibody titers determined by these two assays were well correlated (R(2)=0.7689). These results suggest that the pCNA can serve as a rapid and objective procedure for the measurement of neutralizing antibodies against CV-A16.
authors.affiliation: College of Life Science, Jilin University, Changchun 130012, China. jinjunjlu@126.com
authors.forename: Jun
authors.initials: J
authors.lastname: Jin
authors.forename: Hongxia
authors.initials: H
authors.lastname: Ma
authors.forename: Lin
authors.initials: L
authors.lastname: Xu
authors.forename: Dong
authors.initials: D
authors.lastname: An
authors.forename: Shiyang
authors.initials: S
authors.lastname: Sun
authors.forename: Xueyong
authors.initials: X
authors.lastname: Huang
authors.forename: Wei
authors.initials: W
authors.lastname: Kong
authors.forename: Chunlai
authors.initials: C
authors.lastname: Jiang
mesh_terms: D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral; D045744:Cell Line, Tumor; D002675:Child, Preschool; D019411:Clinical Laboratory Techniques / Q000379:methods*; D004770:Enterovirus / Q000276:immunology*; D017930:Genes, Reporter; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D007223:Infant; D008156:Luciferases / Q000032:analysis; D009500:Neutralization Tests / Q000379:methods*; D044967:Serum / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000956:Antigens, Viral; D008156:Luciferases