Suwimonteerabutr Junpen, Chaicumpa Wanpen, Saengjaruk Patcharin, Tapchaisri Pramuan, Chongsa-nguan Manas, Kalambaheti Thareerat, Ramasoota Pongrama, Sakolvaree Yuwaporn, Virakul Prachin
Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Am J Vet Res. 2005 May;66(5):762-6. doi: 10.2460/ajvr.2005.66.762.
To evaluate the efficacy of a novel monoclonal antibody (MAb)-based dot-blot ELISA for detection of Leptospira antigens in urine samples of cattle.
Blood and urine samples of 45 test cattle from 5 farms in Chonburi province and 20 control cattle from 2 farms in Khon Kaen province in Thailand.
Blood and urine samples were assayed (microscopic agglutination test and urine antigen test) for Leptospira infection by use of an MAb-based dot-blot ELISA, and results for the ELISA were compared with those for dark-field microscopy (DFM), microbial culture, and a polymerase chain reaction (PCR) assay.
All urine samples with positive results for DFM, microbial culture, PCR assay, or > 1 of these tests also had positive results when tested by use of the MAb-based dot-blot ELISA, except for 1 sample that had positive results only for the PCR assay. Detection limits of the dot-blot ELISA were 10(3) leptospires/mL of urine and 9.3 ng of Leptospira homogenate. Comparison revealed that the diagnostic sensitivity, specificity, efficacy (accuracy), positive predictive value, and negative predictive value for the ELISA were in agreement with results for DFM (100%, 72.72%, 80%, 57.14%, and 100%, respectively), microbial culture (100%, 61.54%, 66.62%, 28.57%, and 100%, respectively), and PCR assay (95.45%, 100%, 91.77%, 100%, and 95.83%, respectively).
The MAb-based dot-blot ELISA is suitable as a tool for detecting leptospires in urine samples of cattle.
评估一种基于新型单克隆抗体（MAb）的斑点印迹酶联免疫吸附测定法（ELISA）检测牛尿液样本中钩端螺旋体抗原的效果。
来自泰国春武里府5个农场的45头试验牛以及孔敬府2个农场的20头对照牛的血液和尿液样本。
使用基于MAb的斑点印迹ELISA对血液和尿液样本进行钩端螺旋体感染检测（显微镜凝集试验和尿液抗原检测），并将ELISA结果与暗视野显微镜检查（DFM）、微生物培养及聚合酶链反应（PCR）检测结果进行比较。
所有经DFM、微生物培养、PCR检测或以上多种检测呈阳性的尿液样本，在用基于MAb的斑点印迹ELISA检测时也呈阳性，但有1个样本仅PCR检测呈阳性。斑点印迹ELISA的检测限为每毫升尿液10³个钩端螺旋体和9.3纳克钩端螺旋体匀浆。比较结果显示，ELISA的诊断敏感性、特异性、效能（准确性）、阳性预测值和阴性预测值与DFM结果（分别为100%、72.72%、80%、57.14%和100%）、微生物培养结果（分别为100%、61.54%、66.62%、28.57%和100%）以及PCR检测结果（分别为95.45%、100%、91.77%、100%和95.83%）相符。
基于MAb的斑点印迹ELISA适用于检测牛尿液样本中的钩端螺旋体。
