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基于重组LigB抗原的间接ELISA和乳胶凝集试验用于印度牛钩端螺旋体病血清学诊断的评估

Evaluation of recombinant LigB antigen-based indirect ELISA and latex agglutination test for the serodiagnosis of bovine leptospirosis in India.

作者信息

Deneke Yosef, Sabarinath T, Gogia Neha, Lalsiamthara Jonathan, Viswas K N, Chaudhuri Pallab

机构信息

Genetic Engineering of Bacteria Lab., Division of Bacteriology & Mycology, Indian Veterinary Research Institute, Izatnagar 243122, India.

Leptospira Laboratory, Division of Bacteriology & Mycology, Indian Veterinary Research Institute, Izatnagar 243122, India.

出版信息

Mol Cell Probes. 2014 Aug;28(4):141-6. doi: 10.1016/j.mcp.2014.01.001. Epub 2014 Jan 18.

Abstract

Leptospirosis is a zoonotic disease caused by pathogenic spirochetes of the genus Leptospira, causing febrile infection characterized by multi-organ failure in humans and animals. Leptospiral Ig-like protein B (LigB) is a surface-expressed antigen that mediates host cell invasion or attachment. In this study, N-terminal conserved region of LigB protein (46 kDa) was evaluated for its diagnostic potential to detect anti-leptospiral antibodies in the sera of various animal species. Dot blot analysis revealed immunoreactivity of Leptospira-positive sera of cattle, buffalo, dog, sheep and goat to purified LigB protein. We have analyzed 1126 bovine serum samples, collected from Northern and Eastern part of India, by microscopic agglutination test (MAT) and recombinant LigB (rLigB) based ELISA and latex agglutination test (LAT). The sensitivity of rLigB based ELISA for 554 MAT positive sera was 96.9% and the specificity with 572 MAT negative sera was 91.08% whereas LAT showed sensitivity and specificity of 93.68% and 92.31%, respectively. Kappa values of 0.879 and 0.860 for recombinant antigen based ELISA and LAT indicate excellent agreement with the gold standard serological test, MAT, for the detection of anti-leptospiral antibodies in sera. Further, LAT based on rLigB antigen is a simple and rapid test, suitable for serodiagnosis of leptospirosis under field conditions, owing to its portability and longer shelf life.

摘要

钩端螺旋体病是一种由钩端螺旋体属致病性螺旋体引起的人畜共患病,可导致人和动物出现以多器官功能衰竭为特征的发热性感染。钩端螺旋体免疫球蛋白样蛋白B(LigB)是一种表面表达抗原,介导宿主细胞侵袭或附着。在本研究中,评估了LigB蛋白(46 kDa)的N端保守区域在检测各种动物血清中抗钩端螺旋体抗体方面的诊断潜力。斑点印迹分析显示,牛、水牛、狗、绵羊和山羊的钩端螺旋体阳性血清与纯化的LigB蛋白具有免疫反应性。我们通过显微镜凝集试验(MAT)、基于重组LigB(rLigB)的酶联免疫吸附测定(ELISA)和乳胶凝集试验(LAT),分析了从印度北部和东部收集的1126份牛血清样本。基于rLigB的ELISA对554份MAT阳性血清的敏感性为96.9%,对572份MAT阴性血清的特异性为91.08%,而LAT的敏感性和特异性分别为93.68%和92.31%。基于重组抗原的ELISA和LAT的kappa值分别为0.879和0.860,表明在检测血清中抗钩端螺旋体抗体方面,与金标准血清学检测MAT具有极好的一致性。此外,基于rLigB抗原的LAT是一种简单快速的检测方法,由于其便携性和较长的保质期,适用于现场条件下钩端螺旋体病的血清学诊断。

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