Safe and Objective Assay of Enterovirus 71 Neutralizing Antibodies via Pseudovirus

作     者：JIN Jun1, XU Lin1, GUO Shi-jie2, SUN Shi-yang1, ZHANG Shu1, ZHU Chang-lin3, KONG Wei1 and JIANG Chun-lai1 1. National Engineering Laboratory for AIDS Vaccine, College of Life Science, Jilin University, Changchun 130012, P. R. China 2. Department of Pediatrics, the First Hospital of Jilin University, Changchun 130021, P. R. China 3. Changchun Baike Biotechnology Co., Changchun 130012, P. R. China 

作者机构：National Engineering Laboratory for AIDS Vaccine College of Life Science Jilin University Changchun 130012 Department of Pediatrics the First Hospital of Jilin University Changchun 130021 Changchun Baike Biotechnology Co. Changchun 130012 

出 版 物：《Chemical Research in Chinese Universities》 (高等学校化学研究（英文版）)

年 卷 期：2012年第28卷第1期

页      面：91-95页

核心收录：

学科分类：1010[医学-医学技术（可授医学、理学学位）] 0703[理学-化学] 10[医学] 

基　　金：Supported by the National Natural Science Foundation of China(No.20872048) 

主　　题：Enterovirus 71(EV71) Pseudovirus Luciferase Neutralizing antibody assay 

摘      要：Current serum neutralization assays based on the inhibition of the cytopathic effect（Nt-CPE） need to ma nipulate live viruses, which are time-consuming, labor-intensive, and have the potential exposure to infectious agents, so a safe and objective assay via pseudovirus for the fast and efficient detection of enterovirus 71（EV71） neutralizing antibodies was developed. First, we generated EV71 pseudovirus containing firefly luciferase gene in place of the capsid gene P1 in EV71 genome. Vero cells infected with 200 CCID50（50% cell culture infective dose） of EV71 pseudovirus for 24 h were found to have the best performance. Seval sera were measured by EV71 pseudoparticle neutralization assay（Nt-PPN） and the conventional serological method Nt-CPE. Neutralizing antibody titers measured by Nt-PPN and those obtained by Nt-CPE demonstrate a high correlation between the two methods. Overall, the PPN assay represents a valid alternative to conventional serological methods for the evaluation of EV71 neutralizing anti bodies. This method can be used for detecting neutralizing antibodies of other picornaviruses, such as hepatitis A vi rus（HAV） and coxsackievirus 16（CVA16）, and make it possible to determine whether there is cross-reactivity be tween EV71 and CVA16.