Bastos Carla Resende, Mathias Luis Antonio, Jusi Márcia Mariza Gomes, Santos Renata Ferreira Dos, Silva Glaucenyra Cecília Pinheiro da, André Marcos Rogério, Machado Rosangela Zacarias, Bürger Karina Paes
São Paulo State University (UNESP), School of Agricultural and Veterinarian Sciences, Jaboticabal, Department of Preventive Veterinary Medicine and Animal Reproduction, Jaboticabal, SP, Brazil.
São Paulo State University (UNESP), School of Agricultural and Veterinarian Sciences, Jaboticabal, Department of Preventive Veterinary Medicine and Animal Reproduction, Jaboticabal, SP, Brazil.
Braz J Microbiol. 2018 Jul-Sep;49(3):564-568. doi: 10.1016/j.bjm.2017.10.002. Epub 2018 Feb 13.
The objective of this study was to standardize and validate the dot-blot test for the serological diagnosis of bovine brucellosis, compare the results with those found in the 2-mercaptoethanol (2-ME) and complement fixation test (CF), and estimate the relative sensitivity and specificity of the dot-blot compared to these tests. Fifty bovine blood serum samples were used for the test standardization, and 1315 samples were used for evaluation and comparison between the tests; the results were compared using the Kappa indicator. At the end of standardization, it was established as optimal for the antigen obtained from Brucella abortus B19 after passing through a microorganism rupture process, the blood serum samples diluted at 1:100, and the conjugate at 1:30,000. The comparison of the dot-blot results with 2-ME showed Kappa index of 0.9939, sensitivity of 99.48%, and specificity 99.91%, with CF, Kappa index of 0.8226, sensitivity 100% and specificity 95.32%. Using the combination of the test results 2-ME and CF to establish the true condition of the animal, the dot-blot showed relative sensitivity of 100%, and relative specificity of 99.91%. The evaluated test proved to be effective and reliable, besides being easy to handle and interpret the results.
本研究的目的是对用于牛布鲁氏菌病血清学诊断的斑点印迹试验进行标准化和验证,将结果与在2-巯基乙醇(2-ME)和补体结合试验(CF)中得到的结果进行比较,并评估斑点印迹试验相对于这些试验的相对敏感性和特异性。50份牛血清样本用于试验标准化,1315份样本用于试验间的评估和比较;使用Kappa指标对结果进行比较。标准化结束时,确定经过微生物裂解过程后从流产布鲁氏菌B19获得的抗原、稀释至1:100的血清样本以及稀释至1:30000的结合物为最佳条件。斑点印迹结果与2-ME的比较显示Kappa指数为0.9939,敏感性为99.48%,特异性为99.91%;与CF的比较显示Kappa指数为0.8226,敏感性为100%,特异性为95.32%。使用2-ME和CF的试验结果组合来确定动物的真实状况,斑点印迹试验显示相对敏感性为100%,相对特异性为99.91%。除了易于操作和解释结果外,所评估的试验被证明是有效且可靠的。