PCR方法检测猕猴奇异变形杆菌

Detection of Proteus mirabilisin Macaca mulattaby PCR method

目的利用PCR检测猕猴奇异变形杆菌,为该病菌的鉴定、临床诊断、治疗及流行病学调查等提供参考。方法从猕猴粪便中分离奇异变形杆菌并进行鉴定,纯培养后提取基因组DNA,经PCR扩增、凝胶电泳检测、基因序列测序后,序列提交BLAST比对分析。用该菌株检验PCR方法的特异性和敏感度,利用PCR方法和传统分离培养法同时对80只猕猴进行检测。结果在分离得到的猕猴奇异变形杆菌中扩增出225bp的基因序列(GenBank登录号为JQ040548,其与GenBank中奇异变形杆菌AM942759的同源性为98.2%),而其它5株非奇异变形杆菌的PCR扩增结果为阴性,表现出极好的特异性。纯化培养的奇异变形杆菌利用PCR方法检测灵敏性,其敏感度达80cfu/mL。在80只猕猴中,共检测出了2份阳性样品,阳性率为2.5%,检测结果与传统培养方法一致。结论该PCR方法能迅速、准确地检测猕猴奇异变形杆菌,在临床具有良好的实用性。

In order to provide data for Proteus mirabilis identification, clinical diagnosis, treatment and epidemiological investigation in Macaca mulatta, the Proteus rnirabilis from Macaca mulatta was detected by PCR method in this study. Pro teus mirabilis was isolated and identified from feces of Macaca mulatta, and the DNA was extracted from cultured strains and then amplified by PCR. The products were analyzed by gel electrophoresis and then sequencing. The sequence of Proteus mira- bilis was identified by BLAST. The specificity and sensitivity of PCR were assessed. Eighty samples of Macaca mulatta were screened for Proteus mirabilis by PCR method and traditional method of isolation and identification simultaneously. Results showed that the sequence （225 bp） of Proteus mirabilis was amplified from Macaca rnulatta and submitted to GenBank （No. JQ040548） ; homology analysis showed that the strain shared 98.2 % similarity with the Proteus mirabilis （AM942759）. The specificity was identified by negative amplification of other 5 nomProteus znirabilis strains. The sensitivity of PCR was 80 cfu/mL. The PCR method was comparable with traditional method of isolation and identification as the positive rate of Pro- teus mirabilis in Macaca mulatta detected by that was 2.5% （2/80）. The PCR method was useful for its rapid and accurate de- tection of Proteus mirabilis in Macaca mulatta, which would be applicable in clinic.