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首页> 外文期刊>Folia microbiologica >Rapid and simultaneous detection of Salmonella spp., Escherichia coli O157, and Listeria monocytogenes by magnetic capture hybridization and multiplex real-time PCR
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Rapid and simultaneous detection of Salmonella spp., Escherichia coli O157, and Listeria monocytogenes by magnetic capture hybridization and multiplex real-time PCR

机译：磁捕获杂交和多重实时PCR快速和同时检测沙门氏菌SPP。，大肠杆菌o157，大肠杆菌O157和Histeria单核细胞增生

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摘要

The application of rapid, specific, and sensitive methods for pathogen detection and quantification is very advantageous in diagnosis of human pathogens in several applications, including food analysis. The aim of this study was the evaluation of a method for the multiplexed detection and quantification of three significant foodborne pathogenic species (Escherichia coli O157, Salmonella spp., and Listeria monocytogenes). The assay combines specific DNA extraction by multiplex magnetic capture hybridization (mMCH) with multiplex real-time PCR. The amplification assay showed linearity in the range 10(6)-10 genomic units (GU)/PCR for each co-amplified species. The sensitivity corresponded to 1 GU/PCR for E. coli O157 and L. monocytogenes, and 10 GU/PCR for Salmonella spp. The immobilization process and the hybrid capture of the MCH showed good efficiency and reproducibility for all targets, allowing the combination in equal amounts of the different nanoparticle types in mMCH. MCH and mMCH efficiencies were similar. The detection limit of the method was 10 CFU in samples with individual pathogens and 10(2) CFU in samples with combination of the three pathogens in unequal amounts (amount's differences of 2 or 3 log). In conclusion, this multiplex molecular platform can be applied to determine the presence of target species in food samples after culture enrichment. In this way, this method could be a time-saving and sensitive tool to be used in routine diagnosis.
机译：用于病原体检测和定量的快速，特异性和敏感方法的应用在若干应用中的人类病原体的诊断非常有利，包括食物分析。该研究的目的是评价用于多重检测和定量三种显着的食源性致病物种（大肠杆菌O157，沙门氏菌SPP的方法的方法。该测定用多重实时PCR通过多重磁捕获杂交（MMCH）结合了特异性DNA提取。扩增测定显示为每个共同扩增物种的10（6）-10基因组单元（GU）/ PCR的范围内的线性。对大肠杆菌O157和L.单核细胞生成的1GU / PCR的敏感性对应于1GU / PCR，以及10μmSspp的10Gu / PCR。固定化方法和MCH的杂化捕获显示出所有靶标的良好效率和再现性，允许在MMCH中相等的不同纳米颗粒类型的组合。 MCH和MMCH效率相似。该方法的检测限为10个CFU，其在具有单个病原体和10（2）CFU中的样品中的样品，其中三种病原体的组合在不等量（量的2或3个原点的差异）。总之，该多重分子平台可用于确定培养物富集后食品样品中靶种类的存在。通过这种方式，这种方法可以是常规诊断中使用的节省时间和敏感的工具。

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