A loop-mediated isothermal amplification method targets the phoP gene for the detection of Salmonella in food samples

被引：70

作者：

Li, Xuefei ^{[1
]}

Zhang, Shu ^{[1
]}

Zhang, Hongwei ^{[2
]}

Zhang, Lihuai ^{[2
]}

Tao, Haitao ^{[1
]}

Yu, Jia ^{[3
]}

Zheng, Wenjie ^{[2
]}

Liu, Chenghu ^{[1
]}

Lue, Dan ^{[1
]}

Xiang, Rong ^{[1
]}

Liu, Yin ^{[1
]}

机构：

[1] Nankai Univ, Sch Med, Tianjin 30071, Peoples R China

[2] Tianjin Entry Exit Inspect & Quarantine Bur, Tianjin 300457, Peoples R China

[3] Nankai Univ, Dept Biochem & Mol Biol, Coll Life Sci, Tianjin 300071, Peoples R China

来源：

INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY | 2009年 / 133卷 / 03期

关键词：

Loop-mediated isothermal amplification assay; Salmonella; phoP; Food-borne pathogen; Assay; REAL-TIME PCR; RAPID DETECTION; SEQUENCE;

D O I：

10.1016/j.ijfoodmicro.2009.05.027

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

A loop-mediated isothermal amplification (LAMP) assay was developed and validated for the specific detection of Salmonella in food. The assay used specifically designed primers to target within the phoP gene and correctly identified all 66 strains of Salmonella and 73 non-Salmonella strains tested. The phoP gene regulates the expression of genes involved in virulence and the survival of Salmonella from destruction by macrophage. The probability of detection was 100% when a Salmonella cell suspension containing 10(1) CFU/ml was used as a template in the LAMP assay. Prior to the LAMP assay, a sample preparation protocol was applied that included a pre-enrichment step in buffered peptone water, followed by extraction and purification of DNA. In this way, 85 various food samples were investigated for Salmonella including minced meat of pig and raw milk. The diagnostic accuracy was shown to be 100% when compared to the traditional culture method. This combination of sample enrichment, and LAMP assay can detect 35 CFU per 250 ml of prepared food samples. The overall analysis time for the LAMP assay method was approximately 24 h. This is in contrast to 5 to 7 days of analysis time required for the traditional culture method. Consequently, the LAMP described here has the potential to become a standardized method for the rapid detection of Salmonella in diagnostic laboratories once further validated by inter-laboratory studies. (C) 2009 Elsevier B.V. All rights reserved.

引用

页码：252 / 258

页数：7

共 33 条

[1]

[Anonymous], 65792003 ISO

[2]

CATHARINA CB, 2007, J CLIN MICROBIOL, V45, P1936

[3] The comparative burden of salmonellosis in the European Union member states, associated and candidate countries [J].