Detection of Salmonella spp. by a loop-mediated isothermal amplification (LAMP) method targeting bcfD gene

被引:46
|
作者
Zhuang, L. [1 ,4 ]
Gong, J. [1 ,2 ]
Li, Q. [3 ]
Zhu, C. [2 ]
Yu, Y. [2 ]
Dou, X. [2 ]
Liu, X. [2 ]
Xu, B. [2 ]
Wang, C. [1 ,4 ]
机构
[1] Yangzhou Univ, Coll Vet Med, Yangzhou 225009, Jiangsu, Peoples R China
[2] Chinese Acad Agr Sci, Poultry Inst, Yangzhou, Jiangsu, Peoples R China
[3] Yangzhou Univ, Coll Biosci & Biotechnol, Yangzhou 225009, Jiangsu, Peoples R China
[4] Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou, Jiangsu, Peoples R China
基金
美国国家科学基金会;
关键词
bcfD gene; detection; loop-mediated isothermal amplification; PCR; Salmonella; RAPID DETECTION; SENSITIVE DETECTION; PCR AMPLIFICATION; TYPHIMURIUM; SEQUENCE; POULTRY;
D O I
10.1111/lam.12328
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In this study, we developed and validated a loop-mediated isothermal amplification (LAMP) assay for Salmonella detection targeting bcfD gene, a conserved fimbrial operon gene existing in Salmonella. The Salmonella LAMP assay we developed successfully amplified 44 Salmonella strains (14 standard strains and 30 clinical isolates), but none of 9 non-Salmonella standard strains (Proteus mirabilis, Listeria monocytogenes, Escherichia coli, Pseudomonas aeruginosa, Shigella flexneri, Shigella sonnei, Klebsiella pneumoniae, Campylobacter jejuni and Vibrio parahemolyticus). The detection limit was 5 CFU of Salmonella pure culture or 200 CFU of artificially spiked faeces per reaction system (equivalent to 5000 CFUg(-1) of faeces), and this method could directly detect Salmonella in chicken faeces free of pre-enrichment in a reaction time of 25min. Our experiments show that the LAMP method we developed is a rapid, sensitive, specific and practical method for Salmonella detection. The Salmonella LAMP assay can potentially serve as new on-site diagnostics in the food and agricultural industries.
引用
收藏
页码:658 / 664
页数:7
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