摘要
根据GenBank数据库单增李斯特菌的O基因序列(AF253320)设计引物,建立单增李斯特菌实时荧光定量PCR检测方法。标准曲线的相关系数为0.996,检出底限约8个细菌/反应,而常规PCR检出底限约60个细菌/反应。比较常规PCR和实时荧光定量PCR对人工污染样品检测,发现实时定量PCR也具有较高的灵敏度;对40份牛奶和40份火腿样品进行检测,阳性率分别为12.5%和10%,与传统细菌分离检测结果完全相符。因此,实时定量PCR检测单增李斯特菌具有快速、灵敏的优点,适宜于食品中单增李斯特菌污染的调查及监测。
According to the O gene（AF253320） of L. monocytogene from GenBank, a pair of primers were designed to establish a real-time quantitative PCR assay. The correlation coefficient of standard curve was 0. 996. The minimal detection limit was 8 bacteria/reaction,while the detection limit is 60 bacteria/reaction by conventional PCR. Compared with the conventional PCR and the real-time quantitative PCR to artificially contaminated milk samples and ham samples, the real-time quantitative PCR is more sensitive than conventional PCR. Detecting 40 milk samples and 40 ham samples by real-time quantitative PCR, the positive rate was 12.5% and 10% respectively and the results was identical with the results by conventional microbiology method. Thus, the real-time quantitative PCR is more rapid, sensitive than other methods in detecting L. monocytogene, and is applicable to investigate and monitor L. Monocytogene contamination in food.
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2007年第5期100-104,共5页
Food and Fermentation Industries
基金
河南省科技攻关计划项目(No.0624430008)
关键词
实时定量PCR
食源性致病菌
单增李斯特菌
real-time quantitative PCR, food-borne pathogenic bacteria, L. monocytogene
