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[非洲猪瘟病毒p35蛋白作为诊断抗原的抗原性比较]

[Comparison of the antigenicity of African swine fever virus p35 protein as diagnostic antigen].

作者信息

Shi Lei, Tian Zhancheng, Yang Jifei, Gao Shandian, Du Junzheng, Zhao Yaru, Liu Zhijie, Guan Guiquan, Liu Guangyuan, Luo Jianxun, Yin Hong

机构信息

State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China.

Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, Jiangsu, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2021 Jan 25;37(1):187-195. doi: 10.13345/j.cjb.200359.

Abstract

In order to screen African swine fever virus (ASFV) diagnostic antigen with the best enzyme linked immunosorbent assay (ELISA) reactivity. By establishing the ELISA method, the diagnostic antigen of ASFV p30 protein expressed by baculovirus-insect cell expression system as reference, we explored the antigenic properties and diagnostic potential of ASFV p35 protein expressed by prokaryotic expression system as a diagnostic antigen. The results of Western blotting and immunofluorescence show that the molecular weight of the recombinant p35 protein and p30 protein obtained was 40 kDa and 30 kDa, respectively, and these two proteins had good immuno-reactivity with ASFV positive serum. Recombinant p30 and p35 proteins were used as diagnostic antigens to establish ELISA, and the sensitivity and repeatability of these methods were tested. The results show that although the detection sensitivity of the p30-ELISA established in this study was higher than that of the p35-ELISA, the sensitivity of p35-ELISA was 95.8%, and variations in intra- and inter-assay repeatability of the two methods were less than 10%. The coincidence rate between the p35-ELISA and the imported kit was 97.2%. Results show that p35-ELISA was sensitive and stable, and could detect specific antibodies against ASFV.

摘要

为筛选出酶联免疫吸附测定(ELISA)反应性最佳的非洲猪瘟病毒(ASFV)诊断抗原。通过建立ELISA方法,以杆状病毒-昆虫细胞表达系统表达的ASFV p30蛋白诊断抗原为参照,探究原核表达系统表达的ASFV p35蛋白作为诊断抗原的抗原特性及诊断潜力。蛋白质免疫印迹法和免疫荧光结果显示,获得的重组p35蛋白和p30蛋白分子量分别为40 kDa和30 kDa,且这两种蛋白与ASFV阳性血清具有良好的免疫反应性。将重组p30和p35蛋白用作诊断抗原建立ELISA,并检测这些方法的灵敏度和重复性。结果显示,虽然本研究建立的p30-ELISA检测灵敏度高于p35-ELISA,但p35-ELISA的灵敏度为95.8%,两种方法的批内和批间重复性变异均小于10%。p35-ELISA与进口试剂盒的符合率为97.2%。结果表明,p35-ELISA灵敏且稳定,可检测出抗ASFV的特异性抗体。

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