重组钩端螺旋体免疫球蛋白样b蛋白抗原对野生动物钩端螺旋体病的血清诊断

Yosef Deneke Belachew, R. Deb
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摘要

钩端螺旋体病是热带和亚热带国家以及温带国家的主要威胁。该病由致病性钩端螺旋体引起,在世界许多国家被认为是一种新出现或再出现的疾病。家畜和野生动物的感染可导致经济损失,并可能向社区传播。本研究采用重组LigB蛋白进行胶乳凝集试验,该蛋白是一种交叉反应的脂蛋白,可检测任何致病性钩端螺旋体血清型引起的急性感染。用于钩端螺旋体病的血清诊断。在P QE30表达载体中,用IPTG诱导含有LigB gee n端区域的重组大肠杆菌M15细胞,获得46KDa 6X His标记的LigB蛋白,用Ni-NTA亲和层析纯化后,吸附在乳胶珠表面,对疑似钩端螺旋体病野生野血清进行乳胶凝集试验。共采集野生动物血清80份,其中从昌迪加尔Chhatbir动物园采集野生猫科动物血清27份(老虎18份、狮子8份、美洲虎1份),从拉贾斯坦邦焦特布尔动物园采集野生动物血清42份(老虎8份、狮子4份、豹子6份、猎豹2份、黑羚1份、水牛12份、动物园工作人员9份)和活体啮齿动物3份,从Bohpal Van Vihar国家公园采集野生动物血清8份(老虎4份、豹子3份、狮子1份)。来自印度哈里亚纳邦Biwani迷你动物园的3份血清样本(2只狮子和1只老虎)。结果表明,基于rLigB的LAT检测结果为阳性,显微凝集试验(MAT)再次证实了这一结果。LAT结果与MAT一致。综上所述,基于rLigB的LAT在实验室和现场条件下检测钩端螺旋体病是一种快速、简便、可靠、灵敏度和特异性高的诊断工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Serodiagnosis of wildlife leptospirosis employing recombinant leptospiral immunoglobulin like b protein antigen
Leptospirosis is a major threat in tropical and subtropical countries as well as temperate countries. The disease is caused by pathogenic Leptospira species and considered to be an emerging or re-emerging disease in many countries of the world. Infection in domestic animals and wildlife can lead to economic loss and pose a potential spread to the communities. In the present study recombinant LigB protein is employed in latex agglutination test, which is a cross reacting lipoprotein able to detect acute infection caused by any pathogenic leptospiral serovars. It was employed for serodiagnosis of leptospirosis. The 46KDa 6X His tagged LigB protein, obtained by IPTG induction of recombinant E. coli M15 cells containing the N-terminal region of LigB gee in P QE30 expression vector, was purified by Ni-NTA affinity chromatography and adsorbed on latex bead surface for performing latex agglutination test against Leptospirosis suspected wildlife field sera. A total of  80 wildlife sera samples were collected, including  27 wild feline sera samples (18 tigers, 8 lions, and 1 jaguar) obtained from Chhatbir zoo, Chandigarh, 42 serasamples ( 8 tigers, 4 lions and 6 leopards, 2 cheethals, 1 black buck, 12 buffaloes and  9 zoo staff) sera and 3 live rodents ) were received  from Jodhpur zoo Rajasthan, 8 sera samples (4 tigers, 3 leopards, 1 lion) sera from Van Vihar National park, Bohpal, Madhya Pradesh and 3 sera samples (2 lions,and 1 tiger) received from Biwani Mini zoo, Haryana, India. The result showed that sera were tested positive by rLigB based LAT, which were reconfirmed using microscopic agglutination test (MAT). The results from LAT were in concordance with MAT. In conclusion, rLigB based LAT is a rapid, pen site, reliable diagnostic tool of high sensitivity and specificity, under laboratory and field conditions, for the detection of Leptospirosis .
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