Prokaryotic Expression of NS3 Protein of Duck tembusu virus and Establishment and Application of Its Indirect ELISA Detection Method
AN Er-Li1,2,3,4, JI Xin-Qin1,2,*, RUAN Yong1,3,4,*, CHEN Ze-Hui1,2,3,4, CAO Wei1,2,3,4, WANG Han-Han1,2,3,4, LUO Xiao-Yu1,2,3,4, LONG Dan-Dan1, CHEN Jia-Qi1,2, YANG Chun-Pei5, WU Zong-Hao5, WANG Li-Juan5, YAO Bi-Qiong5
1 Faculty of Animal Sciences, GuiZhou University, Guiyang 550025, China; 2 Guizhou Institute of Animal Disease, GuiZhou University, Guiyang 550025, China; 3 Key Laboratory of Plateau Mountain Animal Genetics, Breeding and Reproduction, Ministry of Education, Guizhou University, Guiyang 550025, China; 4 Key Laboratory of Animal Genetics, Breeding and Reproduction, Guiyang 550025, China; 5 County Animal Disease Prevention and Control Center, Sansui 556500, China
Abstract：To develop an indirect ELISA method for rapid detection of Duck tembusu virus (DTMUV) antibodies, in this study, the prokaryotic recombinant expression vector pET-28a-NS3 was constructed, and NS3 protein obtained through induced expression was used as the coated antigen. After optimizing a series of conditions, an indirect ELISA method for detecting DTMUV antibodies was established. Antibody analysis was performed on 30 serum samples for challenge test and 150 collected serum samples by the established ELISA method. The results showed that the intra-assay coefficient of variation and the coefficient of variation (CV) of the indirect ELISA method was 1.49%~4.31% and 1.22%~6.07%, respectively, which met the national standard. DTMUV antibody could be specifically detected by this method. And the indirect ELISA method had no cross-reaction with Avian influenza virus (AIV) H5 subtype, AIV H7 subtype, Newcastle disease virus (NDV), Duck Circovirus (DuCV) and Duck hepatitis A virus (DHAV-1) positive sera. The coincidence rate of positive and negative detection numbers of challenge test samples and commercial kit was 100%, and the coincidence rate of positive and negative detection numbers of farm samples and commercial kit was 97.33%. In summary, the DTMUV NS3 protein antibody indirect ELISA method established in this study has good specificity and repeatability, and provides a rapid and effective detection method for the investigation of DTMUV epidemiological situation.
安而立, 嵇辛勤, 阮涌, 陈泽辉, 曹伟, 王晗晗, 罗晓宇, 龙丹丹, 陈佳琪, 杨纯培, 吴宗豪, 王丽娟, 姚碧琼. 鸭坦布苏病毒NS3蛋白原核表达及其间接ELISA检测方法的建立与应用[J]. 农业生物技术学报, 2024, 32(10): 2437-2446.
AN Er-Li, JI Xin-Qin, RUAN Yong, CHEN Ze-Hui, CAO Wei, WANG Han-Han, LUO Xiao-Yu, LONG Dan-Dan, CHEN Jia-Qi, YANG Chun-Pei, WU Zong-Hao, WANG Li-Juan, YAO Bi-Qiong. Prokaryotic Expression of NS3 Protein of Duck tembusu virus and Establishment and Application of Its Indirect ELISA Detection Method. 农业生物技术学报, 2024, 32(10): 2437-2446.
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