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Development of a novel dual priming oligonucleotide system‐based PCR assay for specific detection of Salmonella from food samples
Journal of Food Safety ( IF 1.9 ) Pub Date : 2020-03-10 , DOI: 10.1111/jfs.12789 Dan‐Dan Li 1 , Chun‐Bo Hao 2 , Zhong‐Mei Liu 3 , Sui‐Jia Wang 1 , Yu Wang 4 , Zhe Chao 5 , Shen‐Yang Gao 6 , Si Chen 7
Journal of Food Safety ( IF 1.9 ) Pub Date : 2020-03-10 , DOI: 10.1111/jfs.12789 Dan‐Dan Li 1 , Chun‐Bo Hao 2 , Zhong‐Mei Liu 3 , Sui‐Jia Wang 1 , Yu Wang 4 , Zhe Chao 5 , Shen‐Yang Gao 6 , Si Chen 7
Affiliation
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In this study, a novel dual priming oligonucleotide (DPO) system‐based polymerase chain reaction (PCR; DPO system‐based PCR) assay, which detected the fimY gene of Salmonella, was developed for the fast food testing. The DPO system‐based PCR assay allowed a wide range of annealing temperatures at 48–68°C to efficiently amplify fimY gene with an analytical detection limit of 1.2 × 102 CFU/ml for Salmonella in pure cultures and artificially contaminated food matrix. Significantly, the presence of a bubble‐like polydeoxyinosine (polyI) linker in the DPO system brought an unparalleled high specificity in the identification of target bacteria, and consequently, the false positives and mismatches of PCR process can be eliminated in priming. Applying the DPO system‐based PCR assay to 285 collected food samples revealed that 29 samples were positive in this assay, in accordance with the results of conventional culture‐based method, indicating a potential diagnostic capability. The high specificity of the DPO system‐based PCR indicates its great potential to be a quick, reliable and practical method for the detection of Salmonella in foods.
中文翻译：
开发一种新型的基于双引物寡核苷酸系统的PCR检测试剂盒，用于食品样品中沙门氏菌的特异性检测
在这项研究中，开发了一种新型的基于双引物寡核苷酸（DPO）系统的聚合酶链反应（PCR;基于DPO系统的PCR）检测方法，该检测方法可检测沙门氏菌的fimY基因，用于快餐食品检测。基于DPO系统的PCR测定允许在48-68°C的宽范围内的退火温度下有效扩增fimY基因，沙门氏菌的分析检出限为1.2×10 2 CFU / ml纯文化和人工污染的食物基质中。值得注意的是，DPO系统中气泡状的聚脱氧肌苷（polyI）接头的存在为靶细菌的鉴定带来了无与伦比的高特异性，因此，在引物中可以消除PCR过程的假阳性和错配。将DPO系统基于PCR的检测方法应用于285份收集的食物样品中，表明29种样品在此检测方法中呈阳性，这与基于传统培养方法的结果一致，表明具有潜在的诊断能力。基于DPO系统的PCR的高特异性表明它有潜力成为一种快速，可靠和实用的食品中沙门氏菌检测方法。
更新日期：2020-03-10
中文翻译：
开发一种新型的基于双引物寡核苷酸系统的PCR检测试剂盒，用于食品样品中沙门氏菌的特异性检测
在这项研究中，开发了一种新型的基于双引物寡核苷酸（DPO）系统的聚合酶链反应（PCR;基于DPO系统的PCR）检测方法，该检测方法可检测沙门氏菌的fimY基因，用于快餐食品检测。基于DPO系统的PCR测定允许在48-68°C的宽范围内的退火温度下有效扩增fimY基因，沙门氏菌的分析检出限为1.2×10 2 CFU / ml纯文化和人工污染的食物基质中。值得注意的是，DPO系统中气泡状的聚脱氧肌苷（polyI）接头的存在为靶细菌的鉴定带来了无与伦比的高特异性，因此，在引物中可以消除PCR过程的假阳性和错配。将DPO系统基于PCR的检测方法应用于285份收集的食物样品中，表明29种样品在此检测方法中呈阳性，这与基于传统培养方法的结果一致，表明具有潜在的诊断能力。基于DPO系统的PCR的高特异性表明它有潜力成为一种快速，可靠和实用的食品中沙门氏菌检测方法。
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