Serological Analysis by Enzyme-Linked Immunosorbent Assay Using Recombinant Antigen LipL32 for the Diagnosis of Swine Leptospirosis
Serological Analysis by Enzyme-Linked Immunosorbent Assay Using Recombinant Antigen LipL32 for the Diagnosis of Swine Leptospirosis
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DOI：
10.1007/s00284-012-0237-x
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发表时间：
2013-02-01
影响因子：
2.6
通讯作者：
Dellagostin, Odir A.
中科院分区：
文献类型：
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作者：
Hartleben, Claudia P.;Leal, Fernanda M. A.;Dellagostin, Odir A.
钩端螺旋体病是由致病性钩端螺旋体引起的一种重要的全球性人畜共患病。物种猪钩端螺旋体病具有重大的经济影响，因为猪是动物蛋白和副产品的来源。猪钩端螺旋体病的症状是流产、死胎、产下虚弱或生病的仔猪，出现在感染后14-60天。诊断钩端螺旋体病的参考方法是显微镜凝集试验（MAT），其中血清样品与钩端螺旋体血清型的活抗原悬浮液反应。然而，MAT是费力和耗时的诊断程序时，处理大量的样品，因此，正在努力开发新的，敏感的，和具体的诊断测试钩端螺旋体病。本研究采用酶联免疫吸附试验（ELISA）检测钩端螺旋体特异性抗体，并对重组LipL 32进行筛选。用86份经MAT检测的猪血清建立rLipL 32/ELISA。与MAT检测的阳性和阴性血清相比，rLipL 32/ELISA的敏感性为100%，特异性为85.1%，准确性为91.86%。对其他细菌性疾病（地方性肺炎和布鲁氏菌病）未观察到阳性反应。本研究建立的rLipL 32/ELISA检测猪钩端螺旋体抗体方法特异、灵敏、简便，可作为流行病学调查的快速筛查方法。
Leptospirosis is an important global zoonotic disease caused by pathogenic Leptospira spp. species. Swine leptospirosis has a major economic impact because pigs are sources of animal protein and by-products. The signs of swine leptospirosis are abortion, stillbirth, birth of weak or ill piglets, appearing 14-60 days after infection. The reference method for diagnosis of leptospirosis is the microscopic agglutination test (MAT), in which serum samples are reacted with live antigen suspensions of leptospiral serovars. However, MAT is laborious and time consuming as a diagnostic procedure when dealing with a large number of samples; therefore, efforts are being made to develop novel, sensitive, and specific diagnostic tests for leptospirosis. In this study, a recombinant LipL32 based on enzyme-linked immunosorbent assay (rLipL32/ELISA) was evaluated as a screening test for the detection of pathogenic leptospiral-specific antibodies. A total of 86 swine serum samples tested by MAT were used to develop rLipL32/ELISA. Compared to positive and negative sera tested by MAT, rLipL32/ELISA showed 100 % sensitivity, 85.1 % specificity, and 91.86 % accuracy. No positive reaction for other bacterial diseases (enzootic pneumonia and brucellosis) was observed. The rLipL32/ELISA reported in this study is a specific, sensitive, and convenient test for the detection of antibodies against swine leptospiral infection and can be used as a rapid screening test in epidemiological surveys.