Detection of Listeria monocytogenes in food using a combined enrichment/real-time PCR method targeting the prfA gene

被引：176

作者：

Rossmanith, Peter ^{[1
]}

Krassnig, Martina ^{[1
]}

Wagner, Martin ^{[1
]}

Hein, Ingeborg ^{[1
]}

机构：

[1] Univ Vet Med, Inst Milk Hyg Milk Technol & Food Sci, Dept Vet Publ Hlth & Food Sci, A-1210 Vienna, Austria

来源：

RESEARCH IN MICROBIOLOGY | 2006年 / 157卷 / 08期

关键词：

Listeria monocytogenes; real-time PCR; food; enrichment;

D O I：

10.1016/j.resmic.2006.03.003

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

A combined enrichment/real-time PCR method for the detection of Listeria monocytogenes is presented. The method is based on a conventional PCR assay targeting the prfA gene, which has been validated and suggested as an international standard PCR method for identifying L. monocytogenes in food. This real-time PCR assay includes an internal amplification control. Inclusivity and exclusivity were 100% each when testing 100 L. monocytogenes isolates, 30 Listeria spp. isolates other than L. monocytogenes, and 29 non-Listeria isolates. The theoretical detection limit was one copy of the target gene per PCR reaction and the practical detection limit was about 5 copies per PCR. Using the combined enrichment/realtime PCR method, 7.5 CFU/25 ml of artificially contaminated raw milk, and 9, 1, and 1 CFU/15 g of artificially contaminated salmon, pate, and green-veined cheese, respectively, were detected. When analyzing 76 naturally contaminated food samples of various types and comparing the results with the ISO 11290-1 standard method, the relative accuracy was 96%, the relative specificity 100%, and the relative sensitivity, 76.9%. (c) 2006 Elsevier SAS. All rights reserved.

引用

页码：763 / 771

页数：9

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