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旋毛虫重组丝氨酸蛋白酶抑制剂用于猪旋毛虫病免疫诊断的评估

Evaluation of recombinant serine protease inhibitor from Trichinella spiralis for immunodiagnosis of swine trichinosis.

作者信息

Nuamtanong Supaporn, Dekumyoy Paron, Adisakwattana Poom

机构信息

Department of Helminthology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.

出版信息

Southeast Asian J Trop Med Public Health. 2012 Sep;43(5):1094-104.

Abstract

Serine protease inhibitors, known as serpins, are mainly expressed in newborn and early-stage Trichinella spiralis larvae, suggesting that T. spiralis serpin (TsSERP) could be used as antigen for the immunodiagnosis of swine trichinosis. We produced His-tagged recombinant TsSERP (rTsSERP) in Escherichia coli and purified it using a Co(2+)-affinity column. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were performed to determine T. spiralis-infected swine sera samples (n = 5), negative controls (n = 26), and other parasite-infected samples (n = 83). WB showed that T. spiralis-infected sera initially reacted with rTsSERP at day 6 post-infection (dpi), and more strongly in late infection (62 and 84 dpi). However, other parasite-infected sera also elicited cross-reactivity to rTsSERP. On the other hand, indirect ELISA showed that TsSERP was an appropriate antigen for detecting late (> 60 dpi) but not early infection. No cross-reaction was observed with other parasite-infected sera. Sensitivity and specificity of TsSERP-ELISA at 62 dpi was 80% and 100%, respectively, and at 84 dpi 100% and 100%, respectively. These preliminary results show that TsSERP-ELISA method is suitable for the diagnosis of swine trichinosis, and could become the standard test for diagnosis of trichinosis in several hosts, including humans.

摘要

丝氨酸蛋白酶抑制剂,即丝氨酸蛋白酶抑制蛋白,主要在旋毛虫新生幼虫和幼虫早期阶段表达,这表明旋毛虫丝氨酸蛋白酶抑制蛋白(TsSERP)可作为猪旋毛虫病免疫诊断的抗原。我们在大肠杆菌中生产了带有组氨酸标签的重组TsSERP(rTsSERP),并使用钴离子亲和柱进行纯化。采用蛋白质印迹法(WB)和酶联免疫吸附测定法(ELISA)对旋毛虫感染猪血清样本(n = 5)、阴性对照(n = 26)以及其他寄生虫感染样本(n = 83)进行检测。WB结果显示,旋毛虫感染血清在感染后第6天(dpi)开始与rTsSERP发生反应,在感染后期(62和84 dpi)反应更强。然而,其他寄生虫感染血清也与rTsSERP产生交叉反应。另一方面,间接ELISA结果表明,TsSERP是检测旋毛虫晚期感染(> 60 dpi)而非早期感染的合适抗原。未观察到与其他寄生虫感染血清的交叉反应。TsSERP-ELISA在62 dpi时的敏感性和特异性分别为80%和100%,在84 dpi时分别为100%和100%。这些初步结果表明,TsSERP-ELISA方法适用于猪旋毛虫病的诊断,并可能成为包括人类在内的多种宿主旋毛虫病诊断的标准检测方法。

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