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用于检测生肉和即食肉类中沙门氏菌的快速实时聚合酶链反应检测法。

Rapid real-time PCR assay for detecting Salmonella in raw and ready-to-eat meats.

作者信息

Patel Jitu R, Bhagwat Arvind A

机构信息

Food Safety Laboratory, Agricultural Research Service, USDA, 10300 Baltimore Avenue, Beltsville, Maryland 20705-2350, USA.

出版信息

Acta Vet Hung. 2008 Dec;56(4):451-8. doi: 10.1556/AVet.56.2008.4.3.

Abstract

A real-time PCR assay was evaluated for the rapid detection (10 h) of Salmonella in meats using molecular beacon probes available as a commercial kit (iQ-Check, Bio-Rad laboratories). Raw (chicken, pork) and ready-to-eat (RTE) meats were artificially contaminated with Salmonella enterica serovar Typhimurium at the estimated level of 2 to 4 cells per 25 g. After 8 h of pre-enrichment in buffered peptone water, a molecular beacon-based PCR assay was performed to detect contamination in raw and RTE meats. The sensitivity and accuracy of the assay were compared with the conventional USDA microbiological procedure. Comparative evaluation of the USDA procedure with the rapid PCR assay for meat samples (n = 63) revealed 1 false negative pork sample with the PCR assay. All uninoculated controls (n = 34) but one sample were negative by both the 10-h PCR assay and the USDA procedure. Developing rapid pathogen detection methods with shorter pre-enrichment times (8-h) and real-time data monitoring capabilities will benefit the industry in preventing recall of contaminated meats by stopping the contaminated products from being introduced into the marketplace.

摘要

使用市售试剂盒(iQ-Check,伯乐公司)中的分子信标探针,对一种用于快速检测(10小时)肉类中沙门氏菌的实时PCR检测方法进行了评估。将生肉(鸡肉、猪肉)和即食(RTE)肉类用肠炎沙门氏菌鼠伤寒血清型以每25克估计2至4个细胞的水平进行人工污染。在缓冲蛋白胨水中预富集8小时后,进行基于分子信标的PCR检测以检测生肉和即食肉类中的污染情况。将该检测方法的灵敏度和准确性与美国农业部传统微生物学方法进行了比较。对63份肉类样品采用美国农业部方法与快速PCR检测方法进行比较评估,结果显示PCR检测有1份猪肉样品出现假阴性。10小时的PCR检测和美国农业部方法对所有未接种对照(n = 34)除1个样品外均呈阴性。开发具有更短预富集时间(8小时)和实时数据监测能力的快速病原体检测方法,将有助于该行业通过阻止受污染肉类进入市场来防止召回受污染肉类。

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