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首页> 外文期刊>Research in Veterinary Science >Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis
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Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

机译：ELISA方法在犬布鲁氏菌引起的犬布鲁氏菌病血清学诊断中的验证

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摘要

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar-gel immunodiffusion (ACID) test for canine brucellosis, were used as the control panel for B. cants infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies
机译：在本研究中，描述了用于犬布鲁氏菌病血清诊断的酶联免疫吸附测定（ELISA）的有效性。通过ELISA和Western blot（WB）对两种不同的抗原提取物进行了比较，这两种提取物是通过热或超声匀浆法从布鲁氏杆菌的野生分离物中获得的微生物抗原得到的。总共使用145种犬血清来确定ELISA的灵敏度，特异性和准确性，方法如下：（1）通过血液培养和PCR确认的34例患有犬双歧杆菌自然感染的动物的血清，以及51份来自健康的血清经琼脂凝胶免疫扩散（ACID）测试对犬布鲁氏菌病结果阴性的犬，被用作B. cants感染的对照组。 （2）研究在巴西同一地理区域与其他常见狗感染发生交叉反应的可能性，从含有钩端螺旋体，埃里希氏犬，犬瘟热病毒（CDV），新孢子虫（Neospora caninum），巴贝斯犬和利什曼原虫（每组10只）均纳入研究。以可溶性细菌提取物（HE抗原）为抗原的ELISA表现出最佳的灵敏度（91.18％），特异性（100％）和准确性（96.47％）。在WB分析中，HE抗原与不同感染犬的血清无交叉反应，而犬双歧杆菌超声处理则具有由这些血清鉴定的各种蛋白条带。用热可溶性犬双歧杆菌抗原进行标准化ELISA的性能表明，该测定法可用作确认该微生物感染的可靠且实用的方法，同时也是血清流行病学研究的工具

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