新等温扩增技术检测小肠结肠炎耶尔森氏菌  

Detection of Yersinia enterocolitica by a New Isothermal Amplification

作　　者：张霞[1] 温华蔚 倪松[1] 王禹[1] 王淞[1] 高琳[1] 刘掘茫 郑文杰[1] 

机构地区：[1]天津出入境检验检疫局,天津300461

出　　处：《食品研究与开发》2015年第23期113-116,共4页Food Research and Development

基　　金：质检总局科研<生态港外来有害动植物疫病及食源性致病菌风险控制措施研究>资助(2014IK222)

摘　　要：应用新型等温扩增检测技术——交叉引物等温扩增结合免疫金标试纸条建立检测小肠结肠炎耶尔森氏菌的方法。针对小肠结肠耶尔森氏菌16-23S rDNA间区序列设计特异性引物及探针,用54株小肠结肠炎耶尔森氏菌及相近株细菌进行特异性试验;通过纯菌液计数、样品中添菌检测进行灵敏度验证;对677份食品用传统生化国标法进行比较检测试验。建立方法具有较好特异性;增菌液检测灵敏度为10^1cfu/mL,当每25g样品中有10^0cfu菌时经增菌步骤后即可检出,样品检测同传统检测结果比较大致相符,没有漏检,假阳性率较低。建立的新型恒温检测方法可用于食品中小肠结肠炎耶尔森氏菌初筛检测。To develop a cross-priming amplification（CPA） combined with immunoblotting analysis method for the detection of Yersinia enterocolitica on food.Specific primers and probe were designed on the basis of six specific sequences in Y.enterocolitica 16S-23 S rDNA internal transcribed spacer.The specificity of the method was evaluated by 54 different bacterial strains.The sensitivity of the method was evaluated by pure bacteria counted and the sample of adding Y.enterocolitica.To detect 677 samples comparing with the biochemical and culture-based assays.All of the Y.enterocolitica strains showed positive results,and the other bacteria gave negative results.The limit of detection of the CPA method was 10^1 cfu/mL for bacteria in pure culture,and10^0 cfu per 25 g of sample with pre-enrichment.The result of sample detection was broadly consistent with the traditional detection,no omission,low false positive rate.This CPA method can be used for the rapid preliminary screening of Y.enterocolitica.

关 键 词：交叉引物等温扩增技术 小肠结肠炎耶尔森氏菌 食品安全检测 

分 类 号：TS207.4[轻工技术与工程—食品科学]