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利用二维电泳结合 LC-MS/MS 技术对旋毛虫和布氏旋毛虫成虫排泄分泌蛋白的首次分析。

The first analysis of Trichinella spiralis and Trichinella britovi adult worm excretory-secretory proteins by two-dimensional electrophoresis coupled with LC-MS/MS.

机构信息

Witold Stefański Institute of Parasitology, Polish Academy of Sciences, Twarda 51/55, 00-818 Warszawa, Poland.

Witold Stefański Institute of Parasitology, Polish Academy of Sciences, Twarda 51/55, 00-818 Warszawa, Poland.

出版信息

Vet Parasitol. 2021 Sep;297:109096. doi: 10.1016/j.vetpar.2020.109096. Epub 2020 May 17.

DOI:10.1016/j.vetpar.2020.109096
PMID:32564938
Abstract

The two most common Trichinella species present in European countries are Trichinella spiralis and Trichinella britovi. The consumption of raw or undercooked meat with invasive larvae results in trichinellosis. Currently, the most commonly used sources for detecting specific anti-Trichinella antibodies is ELISA with the muscle larvae (ML) excretory-secretory (E-S) proteins. However, these serological methods cannot be efficiently applied at early stage of infection. The aim of the current study was to identify the common and species-specific E-S proteins of T. spiralis and T. britovi adult worms which could have potential for accurate diagnosis of Trichinella infection at an early stage of invasion. Different sets of immunoreactive proteins were identified in T. spiralis and T. britovi proteomes by a combination of two-dimensional electrophoresis (2-DE), immunoblot and LC-MS/MS analysis. Polyubiquitin-B, a possible enoyl-CoA hydratase/isomerase YngF or polyubiquitin-like protein was found to be common for both species; gene ontology analysis confirmed its involvement in proteolysis, oxidation-reduction and translation processes, as well as in molecular transport. These molecules, being secreted or excreted at an early stage of parasite development, may play a critical role in the processes occurring during the initial steps of the host invasion and hence be suitable for diagnostic test development.

摘要

在欧洲国家,两种最常见的旋毛虫物种是旋毛虫和布氏旋毛虫。食用带有侵袭性幼虫的生肉或未煮熟的肉会导致旋毛虫病。目前,检测特异性抗旋毛虫抗体最常用的方法是 ELISA 法,使用肌肉幼虫(ML)排泄-分泌(E-S)蛋白。然而,这些血清学方法不能有效地应用于感染的早期阶段。本研究的目的是鉴定旋毛虫和布氏旋毛虫成虫的共同和种特异性 E-S 蛋白,这些蛋白可能具有在侵袭早期准确诊断旋毛虫感染的潜力。通过二维电泳(2-DE)、免疫印迹和 LC-MS/MS 分析相结合,在旋毛虫和布氏旋毛虫的蛋白质组中鉴定出不同的免疫反应性蛋白。多泛素-B,一种可能的烯酰辅酶 A 水合酶/异构酶 YngF 或泛素样蛋白,被发现对两种物种都是共同的;基因本体分析证实其参与了蛋白酶解、氧化还原和翻译过程,以及分子运输。这些分子在寄生虫发育的早期阶段被分泌或排泄,可能在宿主入侵的初始步骤中发生的过程中发挥关键作用,因此适合开发诊断测试。

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