Detection of Salmonella spp. by a loop‐mediated isothermal amplification (LAMP) method targeting bcfD gene. (19th October 2014)

• Record Type: Journal Article
• Authors: Zhuang, L.; Gong, J.; Li, Q.; Zhu, C.; Yu, Y.; Dou, X.; Liu, X.; Xu, B.; Wang, C.
• Is Part Of: Letters in applied microbiology. Volume 59:Number 6(2014:Dec.)
• Page Start: 658
• Page End: 664
• Publication Date: 2014-10-19
• Subjects: Microbiology -- Periodicals; 660.62
• DOI: 10.1111/lam.12328
• Languages: English
• ISSNs: 0266-8254

Abstract

<abstract abstract-type="main" id="lam12328-abs-0001"> <title>Abstract</title> <sec id="lam12328-sec-1001" sec-type="section"> <p>In this study, we developed and validated a loop‐mediated isothermal amplification (LAMP) assay for <italic>Salmonella</italic> detection targeting <italic>bcfD</italic> gene, a conserved fimbrial operon gene existing in <italic>Salmonella</italic>. The <italic>Salmonella </italic>LAMP assay we developed successfully amplified 44 <italic>Salmonella</italic> strains (14 standard strains and 30 clinical isolates), but none of 9 non‐<italic>Salmonella</italic> standard strains (<italic>Proteus mirabilis</italic>, <italic> Listeria monocytogenes</italic>, <italic> Escherichia coli</italic>, <italic> Pseudomonas aeruginosa</italic>, <italic> Shigella flexneri</italic>, <italic> Shigella sonnei</italic>, <italic> Klebsiella pneumoniae</italic>, <italic> Campylobacter jejuni and Vibrio parahemolyticus</italic>). The detection limit was 5 CFU of <italic>Salmonella</italic> pure culture or 200 CFU of artificially spiked faeces per reaction system (equivalent to 5000 CFU g<sup>−1</sup> of faeces), and this method could directly detect <italic>Salmonella</italic> in chicken faeces free of pre‐enrichment in a reaction time of 25 min. Our experiments show that the LAMP method we developed is a rapid, sensitive, specific and practical method for <italic>Salmonella</italic> detection. The <italic>Salmonella </italic>LAMP assay can potentially serve as new on‐site diagnostics in the food and agricultural industries.</p> </sec> <sec id="lam12328-sec-1002" sec-type="section"> <title>Significance and Impact of the Study</title> <p>A loop‐mediated isothermal amplification (LAMP) assay was established to detect <italic>Salmonella</italic> targeting <italic>bcfD</italic> gene, a conserved fimbrial operon gene. The detection limit was 5 CFU of <italic>Salmonella</italic> pure culture or 200 CFU of artificially spiked faeces per reaction system (equivalent to 5000 CFU g<sup>−1</sup> of faeces), and this method could directly detect <italic>Salmonella</italic> in chicken faeces free of pre‐enrichment in a reaction time of 25 min. The <italic>Salmonella</italic> LAMP assay is a rapid, sensitive, specific and practical method for <italic>Salmonella</italic> detection and can potentially serve as new on‐site diagnostics in the food and agricultural industries.</p> </sec> </abstract>