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利用早期感染不同阶段猪血清筛选旋毛虫成虫期早期抗原基因

Screening of early antigen genes of adult-stage Trichinella spiralis using pig serum from different stages of early infection.

作者信息

Liu P, Wu X P, Bai X, Wang X L, Yu L, Rosenthal B, Blaga R, Lacour S, Vallee I, Boireau P, Gherman C, Oltean M, Zhou X N, Wang F, Zhao Y, Liu M Y

机构信息

Key Laboratory for Zoonosis Research, Ministry of Education, Jilin University, Zoonosis Center of National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, China.

出版信息

Vet Parasitol. 2013 May 20;194(2-4):222-5. doi: 10.1016/j.vetpar.2013.02.001. Epub 2013 Feb 9.

Abstract

The goal of this work was to identify novel, early antigens present in Trichinella spiralis. To this end, a cDNA library generated from 3-day old adult worms (Ad3) was immunologically screened using serum from a pig infected with 20,000 muscle larvae. The serum was obtained from multiple, time course bleeds coinciding with early worm development. Seventeen positive clones were isolated using serum obtained at 20 days post infection (dpi). All clones corresponded to one gene that exhibited high sequence identity with the T. spiralis ATP-dependent RNA helicase DDX19B which is involved in parasite growth and development. In addition, nine additional positive clones representing 5 unique genes were identified when the library was screened with 30 dpi serum; four of these five genes displayed high similarity with members of a putative T. spiralis serine protease family known to be involved in host invasion and host-parasite interactions. The remaining gene aligned with the T. spiralis hypothetical ORF 11.30. The identification of these antigens provides potential candidates for the early diagnosis of trichinellosis and for the development of a vaccine against this parasite.

摘要

这项工作的目标是鉴定旋毛虫中存在的新型早期抗原。为此,使用感染了20,000条肌幼虫的猪的血清,对由3日龄成虫(Ad3)构建的cDNA文库进行免疫筛选。该血清取自与蠕虫早期发育同步的多次定时采血样本。使用感染后20天(dpi)获得的血清分离出17个阳性克隆。所有克隆均对应于一个与旋毛虫ATP依赖性RNA解旋酶DDX19B具有高度序列同一性的基因,该酶参与寄生虫的生长和发育。此外,当用30 dpi血清筛选文库时,鉴定出另外9个代表5个独特基因的阳性克隆;这5个基因中的4个与已知参与宿主入侵和宿主-寄生虫相互作用的假定旋毛虫丝氨酸蛋白酶家族成员具有高度相似性。其余基因与旋毛虫假定的开放阅读框11.30对齐。这些抗原的鉴定为旋毛虫病的早期诊断和针对该寄生虫的疫苗开发提供了潜在的候选物。

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