Validation of a Duplex Real-Time PCR for the Detection of Salmonella spp. in Different Food Products

被引：71

作者：

Anderson, Annette ^{[2
]}

Pietsch, Klaus ^{[2
]}

Zucker, Renate ^{[1
]}

Mayr, Anja ^{[1
]}

Mueller-Hohe, Elke ^{[2
]}

Messelhaeusser, Ute ^{[1
]}

Sing, Andreas ^{[1
]}

Busch, Ulrich ^{[1
]}

Huber, Ingrid ^{[1
]}

机构：

[1] Bavarian Hlth & Food Safety Author, D-85764 Oberschleissheim, Germany

[2] CVUA, D-79114 Freiburg, Germany

来源：

FOOD ANALYTICAL METHODS | 2011年 / 4卷 / 03期

关键词：

Salmonella; Real-Time PCR; invA; Internal Amplification Control; Food; POLYMERASE-CHAIN-REACTION; ANTIMICROBIAL ACTIVITY; ESCHERICHIA-COLI; INTERNAL CONTROL; DIAGNOSTIC PCR; AMPLIFICATION; TYPHIMURIUM; BACTERIA; ASSAY; IDENTIFICATION;

D O I：

10.1007/s12161-010-9142-8

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

A 5' nuclease duplex real-time polymerase chain reaction (PCR) assay was developed and validated with various food products for the specific and fast detection of Salmonella spp. in food. The assay used previously published primers in combination with a newly developed probe targeting the invA gene. An internal amplification control, which is coamplified in a duplex PCR, was included in the assay. The analysis of 1,934 natural food samples with real-time PCR and the cultural method in parallel resulted in a relative accuracy of 100% and 99.84% respectively, depending on the enrichment procedure in which buffered peptone water and selective enrichment in Rappaport-Vassiliadis (RV) broth were employed. The duplex real-time PCR assay has proven to be a specific, sensitive and fast screening method for Salmonella spp. in food. The overall analysis time of the PCR method was approximately 28 h, in contrast to 4 to 5 days with conventional Salmonella diagnostics. The developed assay has been shown to be a reliable diagnostic tool for use in routine analysis. It has been validated thoroughly and has become an official method in Germany for the detection of Salmonella spp. in food.

引用

页码：259 / 267

页数：9

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