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用于乳制品细菌学监测中致病微生物检测的聚合酶链反应(PCR)

Polymerase chain reaction (PCR) for detection of pathogenic microorganisms in bacteriological monitoring of dairy products.

作者信息

Allmann M, Höfelein C, Köppel E, Lüthy J, Meyer R, Niederhauser C, Wegmüller B, Candrian U

机构信息

Laboratory of Food Chemistry, University of Berne, Switzerland.

出版信息

Res Microbiol. 1995 Jan;146(1):85-97. doi: 10.1016/0923-2508(96)80273-7.

Abstract

The presence of pathogenic bacteria poses a serious problem in sustaining the safety of dairy products. Microbiological routine controls of these products make use of selective culture techniques. To detect pathogenic species, isolated colonies are characterized by specific metabolic activities and by serotyping. We present an alternative biochemical approach that does not require culture of bacteria. The total bacterial populations of food samples were isolated by centrifugation and analysed by PCRs specific for pathogenic species. A total of 90 raw milk samples and dairy products made from raw milk were screened by this method for the presence of Listeria monocytogenes, Escherichia coli, enterotoxigenic E. coli, Campylobacter jejuni and C. coli. Detection rates were 12/90 (13%) for L. monocytogenes, 41/90 (46%) for E. coli, 18/90 (20%) for enterotoxigenic E. coli producing heat-labile toxin type I or heat-stable toxin type I, and 6/90 (7%) for C. jejuni or C. coli. Except for the use of different amplification primers, this approach is identical for any bacterial species to be detected. Direct PCR analysis of food samples offers rapid screening for the presence of specific bacteria and enables selection of critical samples prior to culture.

摘要

致病细菌的存在对维持乳制品安全构成严重问题。这些产品的微生物常规控制采用选择性培养技术。为了检测致病菌种,分离出的菌落通过特定的代谢活性和血清分型进行鉴定。我们提出了一种无需培养细菌的替代生化方法。通过离心分离食品样本中的总细菌群体,并通过针对致病菌种的聚合酶链反应(PCR)进行分析。用这种方法对总共90份生乳样本和用生乳制成的乳制品进行筛查,以检测单核细胞增生李斯特菌、大肠杆菌、产肠毒素大肠杆菌、空肠弯曲菌和结肠弯曲菌的存在。单核细胞增生李斯特菌的检出率为12/90(13%),大肠杆菌为41/90(46%),产生I型热不稳定毒素或I型热稳定毒素的产肠毒素大肠杆菌为18/90(20%),空肠弯曲菌或结肠弯曲菌为6/90(7%)。除了使用不同的扩增引物外,这种方法对于任何要检测的细菌种类都是相同的。对食品样本进行直接PCR分析可快速筛查特定细菌的存在,并能在培养前选择关键样本。

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