基于TaqMan探针双重荧光PCR检测食品中肠道沙门氏菌和肠炎沙门氏菌

发布时间：2018-02-25 01:09

本文关键词： 肠道沙门氏菌肠炎沙门氏菌 TaqMan探针双重荧光PCR 检测食品　出处：《现代食品科技》2017年01期 　论文类型：期刊论文

【摘要】：建立基于TaqMan探针双重重实时荧光PCR检测肠道沙门氏菌(Salmonella enterica,SP)和肠炎沙门氏菌(Salmonella Enteritidis,SE)的方法。根据SP的aceA基因(Gen Bank:U43344.1)、肠炎沙门氏菌特异序列SEP(GenBank:AF370707.1),分别设计引物和探针,在ace A探针的5′端标记FAM和SEP探针的5′端标记VIC,建立基于TaqMan探针双重荧光PCR检测方法。试验结果,58株29种不同血清型肠道沙门氏菌均扩增出ace A基因扩增曲线,SEP特异性地扩增出15株SE,而28种不同血清型沙门氏菌和17株变形杆菌等阴性对照株扩增结果均为阴性。ace A和SEP的双重荧光PCR扩增效率分别为100%和104%,R2分别为0.999和0.998,最低检测浓度分别达到280 cfu/m L和260 cfu/m L。建立的方法特异性好、灵敏度高,整个试验可在31 h完成,是快速检测SP和SE的有效方法,可用于食品中SP和SE的特异性检测。
[Abstract]:A method was established for the detection of Salmonella enterica) and Salmonella enteritidisse based on TaqMan probes. According to the aceA gene of SP, GenBank: U43344.1, and the specific sequence of Salmonella entericum AF370707.1, the primers and probes were designed, respectively, for the detection of Salmonella entericum and Salmonella enterica. A double fluorescent PCR detection method based on the 5 'end labeling of FAM with ace A probe and the 5' end labeling of SEP probe with SEP probe was established. The results showed that the amplification curve of ace A gene was obtained from 58 strains of 58 strains of Salmonella enteri of different serotypes. Fifteen SES strains were specifically amplified, and 28 strains of salmonella serotype and 17 strains of Proteus negative control strains were amplified by double fluorescence PCR amplification efficiency of negative. Aace A and SEP were 100% and 104R ~ 2 were 0.999 and 0.998, respectively. The low detection concentration reached 280 cfu/m / L and 260 cfu/m / L, respectively. The whole test can be completed in 31 hours. It is an effective method for rapid detection of SP and SE, and can be used for the specific detection of SP and SE in food.
【作者单位】：广东出入境检验检疫局检验检疫技术中心;黄埔出入境检验检疫局;辽宁出入境检验检疫局;
【基金】：广东省科技项目计划(2013B040402004)
【分类号】：R155.5

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