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首页> 外文期刊>African Journal of Bacteriology Research >Detection of Mycobacterium bovis in whey, by multiplex polymerase chain reaction (PCR) and bacteriological culture
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Detection of Mycobacterium bovis in whey, by multiplex polymerase chain reaction (PCR) and bacteriological culture

机译：多重聚合酶链反应（PCR）和细菌培养法检测乳清中的牛分枝杆菌

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The objective of this study was to detect the presence of Mycobacterium bovis (M. bovis) in whey. A total of 233 cow milk samples were analyzed together with 26 tank milk samples that came from dairy herds of several states of the Mexican Republic (Querétaro, San Luis Potosí, Guanajuato, Hidalgo, Coahuila). DNA was obtained from whey and used for polymerase chain reaction-multiplex (PCR-M). Tuberculosis complex was first identified through the detection of gene RD1. Positive samples were subjected to a second PCR-M with the primers for gene RD9 to identify M. bovis. Samples were bacteriologically cultured using conventional techniques for the isolation of mycobacteria. Cohen’s Kappa test (k) and Pearson’s Chi2 were carried out for statistical analysis. ?A 150 bp amplification product of the RD1 region was obtained, which corresponds to the tuberculosis complex, in 34/233 (14.59%) of the individual milk samples and in 4/26 (15.38%) of the tank milk samples. PCR-M with primer RD9, of the 34 individual samples and the 4 tank milk samples, gave an amplification product of 200 pb, which is the expected product for M. bovis. By bacteriological culture, six isolates were obtained; four in individual whey samples and two from tank milk samples, which were then classified by biochemical tests as M. bovis. The concordance between RD1, RD9 PCR-M and bacteriological culture was low, but there was a significant difference between diagnostic techniques with a P = 0.000. The results showed the potential of the PCR-M as a confirmatory test for the diagnosis of tuberculosis in cattle, as well as the advantage of using whey samples, that may be a possible source of infection for the herd and/or humans.
机译：这项研究的目的是检测乳清中牛分枝杆菌（牛分枝杆菌）的存在。共分析了233份牛奶样品，以及来自墨西哥共和国多个州（克雷塔罗，圣路易斯波托西，瓜纳华托州，伊达尔戈，科阿韦拉）的26个罐装牛奶样品。从乳清中获得DNA，并将其用于聚合酶链反应-多重反应（PCR-M）。结核复合体首先通过检测RD1基因来鉴定。阳性样品用基因RD9的引物进行第二次PCR-M鉴定牛分枝杆菌。使用常规技术对样品进行细菌培养以分离分枝杆菌。进行了Cohen的Kappa检验（k）和Pearson的Chi2进行统计分析。在单独的牛奶样品的34/233（14.59％）和在桶装牛奶样品的4/26（15.38％）中获得了RD1区的150 bp扩增产物，它对应于结核病复合体。在34个单独样品和4个罐装牛奶样品中，用引物RD9进行PCR-M，得到的扩增产物为200 pb，这是牛分枝杆菌的预期产物。通过细菌培养，获得了六种分离株。每份乳清样品中有四份，罐装牛奶样品中有两份，然后通过生化测试将其分类为牛分枝杆菌。 RD1，RD9 PCR-M与细菌培养之间的一致性较低，但是诊断技术之间存在显着差异，P = 0.000。结果表明，PCR-M可以作为牛诊断结核病的确证试验，以及使用乳清样本的优势，乳清样本可能是牛群和/或人类的传染源。

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