乳粉中阪崎克罗诺杆菌快速检测方法应用比较  

Comparative Study on Rapid Detection Methods of Cronobacter Sakazakii in Milk Powder

作　　者：王淞[1,2] 张霞[1,2] 王伟[2] 李莉 朱振元[1] 

机构地区：[1]天津科技大学,天津300457 [2]天津出入境检验检疫局,天津300456 [3]青海出入境检验检疫局,青海西宁810099

出　　处：《食品研究与开发》2016年第1期157-161,共5页Food Research and Development

摘　　要：对几种应用较为广泛的阪崎克罗诺杆菌分子生物学检测技术进行比较分析,探求适用于食品实验室检验要求的快速准确的阪崎克罗诺杆菌检测方法。通过DNA灵敏度检测及样品添菌实验比较交叉引物等温扩增法、实时荧光PCR法、环介导等温扩增法的检测灵敏度;通过实际样品检测,以常规培养生化鉴定为基准方法,进行灵敏度、特异性、假阳性率、假阴性率及相对准确度等5方面比较分析。DNA灵敏度检测显示实时荧光PCR法最灵敏,检测低限在0.9 fg/test;样品添菌实验,3种方法检测低限均可达100 cfu/100 g;与基准方法比较,3种方法灵敏度均达到100%,假阴性率0%,特异性在98.8%～99.0%之间,假阳性率1.0%～1.2%之间,相对准确度98.8%～99.0%之间,均能满足食品实验室检测要求。3种分子生物学方法检测时间短,同基准方法比较,检测结果大致相符,无漏检情况,有假阳性结果,可用于日常阪崎克罗诺杆菌初筛检测。Analyze several widely-applied molecular biological detection techniques of Cronobacter sakazakii,to find the rapid and exact detection method of Cronobacter sakazakii that will meet the food laboratory testing requirements. Compare the detection sensitivity of the methods like cross priming amplification method（CPA）,real-time fluorescent PCR method, loop-mediated isothermal amplification method（LAMP） by the DNA detection sensitivity and sample added bacteria experiment; through the sample testing, compare the sensitivity, specificity, false positive rate, false negative rate and the relative accuracy based on conventional cultivation and biochemical identification method. DNA detection sensitivity showed real-time fluorescent PCR method was the most sensitive, low detection limit was 0.9 fg/test; in the sample add bacteria experiment, three methods to detect the low limit of up to 100 cfu/100 g; sensitivity compared with reference methods, three methods were 100 %, false negative rate of 0 %, the specificity was 98.8 %-99.0 %, false positive rate was1.0 %-1.2 %, relative accuracy of 98.8 %-99.0 %, all of which can meet the demand of food laboratory testing.Compared to the benchmark method, the testing time of three kinds of molecular biology method testing was shorter, the test results were nearly accordance with the traditional standard detection method, no false negative happened, false positive hardly happened. It presented that the three methods can be used for daily Cronobacter sakazakii early screening test.

关 键 词：乳粉 阪崎克罗诺杆菌 交叉引物等温扩增 比较 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]