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鉴定伊氏锥虫免疫优势抗原,用于检测实验感染马属动物的慢性锥虫病。

Identification of immuno-dominant antigens of Trypanosoma evansi for detection of chronic trypanosomosis using experimentally infected equines.

机构信息

National Research Centre on Equines, Sirsa Road, Hisar 125001, Haryana, India.

出版信息

Res Vet Sci. 2013 Oct;95(2):522-8. doi: 10.1016/j.rvsc.2013.04.030. Epub 2013 May 25.

Abstract

Trypanosoma evansi is the most extensively distributed trypanosome responsible for disease called surra in livestock in many countries including frequent outbreaks in India. The prevalence of this disease is most commonly reported by standard parasitological detection methods (SPDM); however, antibody ELISA is being in practice by locally produced whole cell lysate (WCL) antigens in many countries. In the present investigation, we attempted to identify and purify immuno dominant, infection specific trypanosome antigens from T. evansi proteome using experimentally infected equine serum by immuno blot. Three immuno dominant clusters of proteins i.e. 62-66 kDa, 52-55 kDa and 41-43 kDa were identified based on their consistent reactivity with donkey sequential serum experimentally infected T. evansi up to 280 days post infection (dpi). The protein cluster of 62-66 kDa was purified in bulk in native form and comparatively evaluated with whole cell lysate antigen (WCL). ELISA and immuno blot showed that polypeptide of this cluster is 100% sensitive in detection of early and chronic infection. Further, this protein cluster was also found immuno reactive against hyper immune serum raised against predominantly 66 kDa exo antigen, revealed that this is a common immunodominant moieties in proteome and secretome of T. evansi.

摘要

伊氏锥虫是分布最广泛的锥虫,可导致许多国家(包括印度)的家畜出现苏拉病。该病的流行情况通常通过标准寄生虫检测方法(SPDM)报告;然而,在许多国家,酶联免疫吸附试验(ELISA)正在使用本地生产的全细胞裂解物(WCL)抗原进行。在本研究中,我们试图通过免疫印迹,使用实验感染马的血清从伊氏锥虫蛋白质组中鉴定和纯化免疫显性、感染特异性锥虫抗原。根据其与驴顺序血清的持续反应性,基于其与驴顺序血清的持续反应性,从蛋白质组中鉴定出三个免疫显性蛋白簇,即 62-66 kDa、52-55 kDa 和 41-43 kDa,直到感染后 280 天(dpi)。以天然形式大量纯化 62-66 kDa 的蛋白簇,并与全细胞裂解物抗原(WCL)进行比较评估。ELISA 和免疫印迹显示,该蛋白簇的多肽在检测早期和慢性感染时具有 100%的敏感性。此外,该蛋白簇还对针对主要 66 kDa 外抗原的高免疫血清呈免疫反应性,表明这是 T. evansi 蛋白质组和分泌组中的共同免疫显性部分。

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