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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Comparison of Real-Time PCR Reverse Transcriptase Real-Time PCR Loop-Mediated Isothermal Amplification and the FDA Conventional Microbiological Method for the Detection of Salmonella spp. in Produce
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Comparison of Real-Time PCR Reverse Transcriptase Real-Time PCR Loop-Mediated Isothermal Amplification and the FDA Conventional Microbiological Method for the Detection of Salmonella spp. in Produce

机译：实时荧光定量PCR逆转录酶实时荧光定量PCR环介导的等温扩增和FDA常规微生物学方法检测沙门氏菌的比较。在生产中

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Contamination of foods, especially produce, with Salmonella spp. is a major concern for public health. Several methods are available for the detection of Salmonella in produce, but their relative efficiency for detecting Salmonella in commonly consumed vegetables, often associated with outbreaks of food poisoning, needs to be confirmed. In this study, the effectiveness of three molecular methods for detection of Salmonella in six produce matrices was evaluated and compared to the FDA microbiological detection method. Samples of cilantro (coriander leaves), lettuce, parsley, spinach, tomato, and jalapeno pepper were inoculated with Salmonella serovars at two different levels (105 and <101 CFU/25 g of produce). The inoculated produce was assayed by the FDA Salmonella culture method (Bacteriological Analytical Manual) and by three molecular methods: quantitative real-time PCR (qPCR), quantitative reverse transcriptase real-time PCR (RT-qPCR), and loop-mediated isothermal amplification (LAMP). Comparable results were obtained by these four methods, which all detected as little as 2 CFU of Salmonella cells/25 g of produce. All control samples (not inoculated) were negative by the four methods. RT-qPCR detects only live Salmonella cells, obviating the danger of false-positive results from nonviable cells. False negatives (inhibition of either qPCR or RT-qPCR) were avoided by the use of either a DNA or an RNA amplification internal control (IAC). Compared to the conventional culture method, the qPCR, RT-qPCR, and LAMP assays allowed faster and equally accurate detection of Salmonella spp. in six high-risk produce commodities.
机译：沙门氏菌污染食品，特别是农产品。是公共卫生的主要问题。有几种方法可用于检测农产品中的沙门氏菌，但需要确认它们在通常食用的蔬菜中通常与食物中毒暴发有关的沙门氏菌检测的相对效率。在这项研究中，评估了三种分子方法在六个农产品基质中检测沙门氏菌的有效性，并与FDA微生物检测方法进行了比较。用两种不同水平的沙门氏菌沙门氏菌接种香菜（香菜叶），生菜，欧芹，菠菜，番茄和墨西哥胡椒的样品（10 5 和<10 1 CFU） / 25克产品）。通过FDA沙门氏菌培养法（细菌学分析手册）和三种分子方法对接种的产品进行测定：定量实时PCR（qPCR），定量逆转录酶实时PCR（RT-qPCR）和环介导的等温扩增（灯）。通过这四种方法获得了可比的结果，全部检测到沙门氏菌细胞/ 25 g产品低至2 CFU。通过四种方法，所有对照样品（未接种）均为阴性。 RT-qPCR仅能检测活的沙门氏菌细胞，从而避免了非存活细胞产生假阳性结果的危险。通过使用DNA或RNA扩增内部对照（IAC）避免了假阴性（抑制qPCR或RT-qPCR）。与常规培养方法相比，qPCR，RT-qPCR和LAMP分析可更快，同样准确地检测沙门氏菌。六种高风险农产品。

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