Abstract：Objective To establish an enzyme-linked immunosorbent assay (ELISA) detection reagent based on Leptospira fusion protein LipL32- LipL41 as a coated antigen.MethodsLeptospira interrogans lipoprotein32 (LipL32) and lipoprotein41 (LipL41) were selected as research targets. The fusion gene lipL32-lipL41 was constructed by using PCR with ligation primers. The recombinant protein was expressed in prokaryotic expression system. The ELISA detection system was established by using the fusion protein as the coated antigen. The sensitivity, specificity and stability of the detection system were evaluated.Results About 1,900 bp lipL32-lipL41 fusion gene fragment was obtained. The molecular weight of the recombinant fusion protein LipL32-LipL41 prokaryotically expressed was about 90 kDa, and an ELISA detection system based on LipL32-LipL41 was established. 77 confirmed serum samples and 85 negative control ones were used for preliminary verification. The sensitivity, specificity, crude consistency and Yorden index of the detection system were 96.1%, 97.6%, 96.9% and 0.937 respectively.Conclusions It is preliminarily verified that the ELISA diagnostic kit based on the recombinant fusion protein LipL32-LipL41 as a coated antigen has high sensitivity, specificity and stability, and can be proceeded with the next verification.
胡玉山, 刘巧谊, 肖丽红, 周勇, 侯水平, 刘俊华. 以重组融合蛋白为基础的钩端螺旋体酶联免疫吸附试验检测试剂盒的初步研制[J]. 实用预防医学, 2019, 26(7): 808-814.
HU Yu-shan, LIU Qiao-yi, XIAO Li-hong, ZHOU Yong, HOU Shui-ping, LIU Jun-hua. Preliminary development of an enzyme-linked immunosorbent assay kit based on recombinant fusion antigen protein for detection of Leptospira. , 2019, 26(7): 808-814.
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2019, Vol. 26 
