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首页> 外文期刊>Archiv fur Lebensmittelhygiene >Comparison of four real-time polymerase chain reaction-based methods for the detection of Salmonella enterica in food
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Comparison of four real-time polymerase chain reaction-based methods for the detection of Salmonella enterica in food

机译：四种基于实时聚合酶链反应的食品中沙门氏菌检测方法的比较

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Four real-time PCR-based methods for the detection of Salmonella (5.) enterica in food, namely,TaqMan Salmonella enterica Detection Kit, iQ-Check Salmonella, BACldent Salmonella spp., and our in-house method (Kuchta et al., 2007), were evaluated. Allmethods produced definitive results on the next day after sample receipt and contained an internal amplification control to monitor potential false negative results. Five naturally contaminated food samples and 20 food samples artificially contaminated with S. enterica at a level of 10~0 cfu / 25 g were analysed by the realtime PCR-based methods in parallel to the standard microbiological method EN ISO 6579. Using the TaqMan kit, positive results (identical to EN ISO 6579) were obtained with all but one food sample (naturally contaminated minced meat), which was attributed to an inappropriate enrichment used with this kit. Using the iQ-Check kit, positive results (identical to EN ISO 6579) were obtained with most food samples, but PCR inhibition was observed with some samples (artificially contaminated cream cakes). This was attributed to a lower efficiency of the lysate purification procedure of the kit with this type of food samples. Using the BAC/denf kit, positive results (identical to EN ISO 6579) were obtained with most food samples, but false negative results were obtained with naturally contaminated black pepper as well as with artificially contaminated red pepper, bread crumbs and with some samples of ice cream.This was attributed to a lower sensitivity of the PCR system, which was probably caused by the degradation of active components in a fully integrated reaction mixture of this kit. Further, PCR inhibition was observed with some samples of cream cakes, which was attributed to the lackof a lysate purification procedure while applying this kit. Our in-house real-time PCR-based method produced positive results (identical to EN ISO 6579) with all food samples.
机译：四种基于PCR的实时检测食品中沙门氏菌（5.）的方法，即TaqMan沙门氏菌检测试剂盒，iQ-Check沙门氏菌，BACldent沙门氏菌属和我们的内部方法（Kuchta等。 （2007年）进行了评估。所有方法在收到样品后的第二天产生确定的结果，并包含内部扩增对照物以监测潜在的假阴性结果。通过基于PCR的实时分析方法与标准微生物学方法EN ISO 6579并行，分析了5种自然污染的食品样品和20种食品样品，这些样品被10.0 cfu / 25 g的肠炎沙门氏菌人工污染。使用TaqMan试剂盒，除一个食品样品（自然污染的肉末）外，所有样品均获得了阳性结果（与EN ISO 6579相同），这归因于该试剂盒使用了不适当的浓缩液。使用iQ-Check试剂盒，大多数食品样品均获得了阳性结果（与EN ISO 6579相同），但某些样品（被人工污染的奶油蛋糕）却被PCR抑制。这归因于这种类型的食物样品的试剂盒的裂解物纯化程序效率较低。使用BAC / denf试剂盒，大多数食品样品均获得了阳性结果（与EN ISO 6579相同），而天然污染的黑胡椒，人工污染的红辣椒，面包屑以及一些食品样品均获得了假阴性结果。冰淇淋的原因是PCR系统灵敏度较低，这可能是由于该试剂盒完全整合的反应混合物中活性成分的降解所致。此外，在某些奶油蛋糕样品中观察到了PCR抑制作用，这归因于使用该试剂盒时缺乏裂解物纯化程序。我们基于内部实时PCR的方法对所有食品样品均产生了积极的结果（与EN ISO 6579相同）。

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