青藏高原牦牛出血性败血症菌落PCR快速诊断技术研究

A species-specific colony PCR method targeted KMT1 gene for diagnosing hemorrhagic septicaemia colonies of yak on Tibet-plateau

为了探索一种快速、特异和敏感的牦牛出血性败血症诊断方法,本研究建立了针对多杀性巴氏杆菌种特异性KMT1基因的菌落PCR方法。结果显示:该方法的阳性检出率达到40.48%(51/126),高于传统细菌分离鉴定方法 28.57%(36/126);其能检出1 pg的DNA,并且特异性好、灵敏度高,对大肠埃希氏菌等3株对照菌均未扩增出253 bp的特异性条带。表明该方法可用于兽医临床和基层进行牦牛出血性败血症的快速诊断,具有较强的临床应用价值。

It is very important to explore a simple, rapid, specific and sensitive diagnosis method for hemorrhagic septicaemia in yak. KMT 1 is an species - specific gene in Pasteurulla multocida （ P. multocida） . A specific colony PCR method targeted to K M T 1 was established for detection of P. multocida in present study. The result showed that the detection rate was 40. 4 8 % （51/126 ） using the PCR method, compared to 28.5 7 % （ 36/126 ） using the traditional method. It was shown that the primer set amplified a 253 bp DNA fragment specifically from P. multocida, but not from other common bacterial. Moreover, results displayed that the sensitivity of the PCR assay was established by the detection of DNA as low as 1pg. The establishment of the specific PCR assay provides a useful tool for the identification and diagnosis of yak infection with P. multocida.