[关键词]
[摘要]
弓形菌是一种新型食源性致病菌,其中以布氏弓形菌污染率最高。本研究采用扩增23S rRNA的PCR法特异性检测布氏弓形菌,方法灵敏度可达103 CFU/mL。2株布氏弓形菌均特异性地扩增出了长度为2061 bp的条带;嗜低温弓形菌、斯氏弓形菌、空肠弯曲菌等共18株不同种类的菌株均无扩增产物出现,表明此PCR法能特异性的将布氏弓形菌鉴定到种一级水平。比较实验结果表明,API CAMPY鉴定试剂盒对于布氏弓形菌鉴定仅能到弓形菌属一级水平,本PCR法能实现布氏弓形菌种一级水平的鉴定,用于布氏弓形菌的检测具有优势。55份动物源性食品样品用Johnson-Murano肉汤增菌后用PCR法进行检测,其中5份样品为布氏弓形菌阳性,阳性检出率为9.1%。上述实验结果表明,本方法特异性强、操作简便,节省了检测时间,可用于动物源性食品中布氏弓形菌的快速检测。
[Key word]
[Abstract]
Arcobacter spp. are newly emerging human foodborne pathogens, of which A. butzleri has the highest contamination rate PCR method amplifying 23S rRNA gene was applied to specifictly detect Arcobacter butzleri in this study, and the sensitivity of the method was 103 CFU/mL. Both of the 2 different reference strains of A. butzleri amplified and formed a band with length of 2061 bp, while A. cryaerophilus, A. skirrowii, Campylobacterjejuni and other different strains, 18 strains in total, didn’t produced any band. The results proved the PCR method had high specificity and could be used to identify A. butzleri at the species level. The comparative experimental results showed the PCR method also had advantage over the API CAMPY test kit, as the latter could only be used to identify A. butzleri at the genus level. 55 animal derived food samples containing the strains were enriched in Johnson-Murano broth and then detected by the PCR method, of which 5 samples were detected positive for A. butzleri, with the positive ratio of 9.1%. It suggested that the method was specific, easy to operate and timesaving, which could be used for rapid detection of A. butzleri in animal derived food.
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[基金项目]
2011年度国家质检总局科研计划项目(2011IK256)