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猪带绦虫囊尾蚴蛋白TsAg5在人类神经囊尾蚴病诊断中具有微弱的类胰蛋白酶活性。

TsAg5, a Taenia solium cysticercus protein with a marginal trypsin-like activity in the diagnosis of human neurocysticercosis.

作者信息

Rueda Analiz, Sifuentes Cecilia, Gilman Robert H, Gutiérrez Andrés H, Piña Ruby, Chile Nancy, Carrasco Sebastián, Larson Sandra, Mayta Holger, Verástegui Manuela, Rodriguez Silvia, Gutiérrez-Correa Marcel, García Héctor H, Sheen Patricia, Zimic Mirko

机构信息

Laboratorio de Bioinformática y Biología Molecular, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias y Filosofía, Universidad Peruana Cayetano Heredia, Lima, Peru.

出版信息

Mol Biochem Parasitol. 2011 Dec;180(2):115-9. doi: 10.1016/j.molbiopara.2011.08.003. Epub 2011 Aug 26.

Abstract

Neurocysticercosis is an endemic parasitic disease caused by Taenia solium larva. Although the mechanism of infection is not completely understood, it is likely driven by proteolytic activity that degrades the intestinal wall to facilitate oncosphere penetration and further infection. We analyzed the publicly available T. solium EST/DNA library and identified two contigs comprising a full-length cDNA fragment very similar to Echinococcus granulosus Ag5 protein. The T. solium cDNA sequence included a proteolytic trypsin-like-domain in the C-terminal region, and a thrombospondin type-1 adherence-domain in the N-terminal region. Both the trypsin-like and adherence domains were expressed independently as recombinant proteins in bacterial systems. TsAg5 showed marginal trypsin-like activity and high sequence similarity to Ag5. The purified antigens were tested in a Western immunoblot assay to diagnose human neurocysticercosis. The sensitivity of the trypsin-like-domain was 96.36% in patients infected with extraparenchymal cysts, 75.44% in patients infected with multiple cysts, and 39.62% in patients with a single cyst. Specificity was 76.70%. The thrombospondin type-1 adherence-domain was not specific for neurocysticercosis.

摘要

神经囊尾蚴病是一种由猪带绦虫幼虫引起的地方性寄生虫病。尽管感染机制尚未完全明确,但可能是由蛋白水解活性驱动,这种活性会降解肠壁以促进六钩蚴穿透并进一步感染。我们分析了公开可用的猪带绦虫EST/DNA文库,并鉴定出两个重叠群,其包含一个与细粒棘球绦虫Ag5蛋白非常相似的全长cDNA片段。猪带绦虫cDNA序列在C端区域包含一个蛋白水解性胰蛋白酶样结构域,在N端区域包含一个血小板反应蛋白1型黏附结构域。胰蛋白酶样结构域和黏附结构域均在细菌系统中作为重组蛋白独立表达。猪带绦虫Ag5(TsAg5)显示出微弱的胰蛋白酶样活性,且与Ag5具有高度序列相似性。纯化后的抗原通过蛋白质免疫印迹法检测以诊断人类神经囊尾蚴病。胰蛋白酶样结构域在脑实质外囊肿感染患者中的敏感性为96.36%,在多发囊肿感染患者中为75.44%,在单发囊肿患者中为39.62%。特异性为76.

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