Detection of Brucella infection in dairy buffaloes using conventional and molecular methods

S. Hazem, Rania I. Ismail, Hend I. Elsharkawy, Eman I. M. Beleta, Abeer A. E. Mohamed
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Abstract

T his study was conducted on different dairy buffalo herds. A total of 316 buffaloes including 290 buffaloes suffering from reproductive disorders and 26 apparently healthy buffaloes in different rural areas belonging to the Al-Qalyubia Governorate were employed in this study. Epidemiological data revealed that these animals had no history of vaccination against Brucellosis. Blood sera of these animals were subjected to serological assessments using the Rose Bengal plate test (RBPT), Competitive En-zyme Linked Immunosorbent Assay (cELISA), complement fixation test (CFT) in addition to the use of real-time PCR ( rtPCR). MRT and bacteriological examination were performed on milk samples. The obtained results showed that the overall sero-prevalences of brucellosis were recorded as 17.41% , 17.09 % , 16.77% , and 17.09% by RBT, c-ELISA, CFT and rtPCR respectively. B. melitensis biovar 3 was isolated from milk samples 47 (14.87%) It was concluded that for the diagnosis of brucellosis in dairy buffaloes, MRT and RBT remain recommended for screening, CFT is ad-vised for confirming infection in individual animals, and cELISA can be used instead of CFT in some cases. Using rtPCR on serum samples should be regarded as an additional diagnostic technique for detecting and identify-ing Brucella infection and should be considered in conjunction with other serological tests that can progress and conquer the limits of different immunoassays. According to bacteriological testing, B. melitensis biovar 3 is still the prevalent Brucella field strain
常规方法与分子方法检测奶牛布鲁氏菌感染
这项研究是在不同的奶牛群中进行的。在这项研究中,共使用了316头水牛,其中包括290头患有生殖障碍的水牛和26头明显健康的水牛,它们生活在属于Al-Qalyubia省的不同农村地区。流行病学资料显示,这些动物没有接种布鲁氏菌病疫苗的历史。采用玫瑰孟加拉板试验(RBPT)、竞争性酶联免疫吸附试验(cELISA)、补体固定试验(CFT)以及实时荧光定量PCR (rtPCR)对这些动物的血清进行血清学评估。对牛奶样品进行MRT和细菌学检查。结果显示,RBT法、c-ELISA法、CFT法和rtPCR法测定的布鲁氏菌病总体血清患病率分别为17.41%、17.09%、16.77%和17.09%。结果表明,对于乳水牛布鲁氏菌病的诊断,建议采用MRT和RBT进行筛查,建议采用CFT进行个体感染的确认,在某些情况下可采用cELISA代替CFT。对血清样本使用rt - pcr应被视为检测和鉴定布鲁氏菌感染的一种额外的诊断技术,并应考虑与其他血清学测试结合使用,这些测试可以取得进展并克服不同免疫测定法的局限性。细菌学检测结果显示,3型布氏菌仍是布鲁氏菌野外流行菌株
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