摘要
目的建立基于新型Taqman荧光探针的荧光实时（Real-time）PCR方法用于空肠弯曲菌的筛选检测与快速鉴定。空肠弯曲菌（Campylobacter jejuni）是近十几年来在世界范围内广泛重视的人畜共患病原菌，可以导致人类的急性肠炎与食物中毒，并能引发格林-巴利综合征等并发症。为更好地利用分子生物学技术对空肠弯曲菌进行快速、准确的基因检测，本研究从样品增菌液与单菌落中提取细菌DNA，建立了针对空肠弯曲菌特异的hipo（hippuricase，马尿酸酶）基因的Real-timePCR方法，用于空肠弯曲菌的快速筛选检测与可疑菌落快速鉴定。试验结果表明，该方法检测细菌的灵敏度为110CFU．人工布菌鸡肉的检测灵敏度为1~10CFU。本研究所建立的空肠弯曲菌荧光实时PCR方法具有准确、可靠、快速的特点。
Campylobacterjejuni is a leading food-borne pathogen of humans,and it can cause severe diarrhea and gastroenteritis. To obtain a rapid and accurate method of detection for C. jejuni by means of molecular biological techniques,a fluorescent real-time PCR assay based on the new type Taqman fluorescence probe was set up to detect the hippuricase （hipO） gene of C.jejun. The DNA used was extracted from the enrichment both cultures and the single bacterial colonys, It was found that the sensitivity of this assay for the detection of C. jejuni was 1-10 CFU. This method appears to be a accurate and highly sensitive and specific assay for the detection of C. jejuni in samples.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2007年第8期785-787,共3页
Chinese Journal of Zoonoses
