title: Efficient and Specific Detection of Salmonella in Food Samples Using a stn-Based Loop-Mediated Isothermal Amplification Method.
abstract: The Salmonella enterotoxin (stn) gene exhibits high homology among S. enterica serovars and S. bongori. A set of 6 specific primers targeting the stn gene were designed for detection of Salmonella spp. using the loop-mediated isothermal amplification (LAMP) method. The primers amplified target sequences in all 102 strains of 87 serovars of Salmonella tested and no products were detected in 57 non-Salmonella strains. The detection limit in pure cultures was 5 fg DNA/reaction when amplified at 65°C for 25 min. The LAMP assay could detect Salmonella in artificially contaminated food samples as low as 220 cells/g of food without a preenrichment step. However, the sensitivity was increased 100-fold (~2 cells/g) following 5 hr preenrichment at 35°C. The LAMP technique, with a preenrichment step for 5 and 16 hr, was shown to give 100% specificity with food samples compared to the reference culture method in which 67 out of 90 food samples gave positive results. Different food matrixes did not interfere with LAMP detection which employed a simple boiling method for DNA template preparation. The results indicate that the LAMP method, targeting the stn gene, has great potential for detection of Salmonella in food samples with both high specificity and high sensitivity.
authors.affiliation: Department of Biotechnology, Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand ; Mahidol University-Osaka University Collaborative Research Center for Bioscience and Biotechnology (MU-OU:CRC), Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand.
authors.forename: Mevaree
authors.initials: M
authors.lastname: Srisawat
authors.affiliation: Department of Biotechnology, Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand ; Mahidol University-Osaka University Collaborative Research Center for Bioscience and Biotechnology (MU-OU:CRC), Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand.
authors.forename: Watanalai
authors.initials: W
authors.lastname: Panbangred
mesh_terms: D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D004768:Enterotoxins / Q000235:genetics; D005506:Food Contamination; D005516:Food Microbiology; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D012475:Salmonella / Q000187:drug effects / Q000235:genetics*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013696:Temperature
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D004768:Enterotoxins