title: Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples.
abstract: We developed and evaluated the specificity and sensitivity of a loop-mediated isothermal amplification (LAMP) method for rapid detection of the food-borne Escherichia coli O157 strains. Six primers, including outer primers, inner primers and loop primers, were specially designed for recognizing eight distinct sequences on three targets, which were rfbE, stx1 and stx2. The detection limits were found to be 100, 100 and 10 fg DNA/tube for rfbE, stx1 and stx2, respectively. Application of LAMP assays were performed on 417 food-borne E. coli strains, the sensitivity of LAMP assays for the rfbE, stx1 and stx2 was 100, 95.3 and 96.3%, and the negative predictive value was 100, 96.7 and 97.1%, respectively; with a 100% specificity and positive predictive value for all three targets.
authors.affiliation: College of Light Industry and Food Sciences, South China University of Technology, 510640 Guangzhou, China.
authors.forename: Xihong
authors.initials: X
authors.lastname: Zhao
authors.forename: Yanmei
authors.initials: Y
authors.lastname: Li
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Lijun
authors.initials: L
authors.lastname: You
authors.forename: Zhenbo
authors.initials: Z
authors.lastname: Xu
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.forename: Xiaowei
authors.initials: X
authors.lastname: He
authors.forename: Yao
authors.initials: Y
authors.lastname: Liu
authors.forename: Jihua
authors.initials: J
authors.lastname: Wang
authors.forename: Liansheng
authors.initials: L
authors.lastname: Yang
mesh_terms: D001681:Biological Assay; D017931:DNA Primers / Q000378:metabolism; D019453:Escherichia coli O157 / Q000235:genetics / Q000302:isolation & purification*; D005504:Food Analysis / Q000379:methods*; D005516:Food Microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D017931:DNA Primers
references.citation: FEMS Microbiol Lett. 2008 Jan;278(2):223-30
references.pmid: 18096018
references.citation: Curr Microbiol. 2008 Sep;57(3):264-8
references.pmid: 18594909
references.citation: J Food Prot. 2005 Oct;68(10):2140-3
references.pmid: 16245720
references.citation: Appl Environ Microbiol. 2001 Jul;67(7):2908-15
references.pmid: 11425701
references.citation: Biochem Biophys Res Commun. 2001 Nov 23;289(1):150-4
references.pmid: 11708792
references.citation: Nucleic Acids Res. 1998 Feb 15;26(4):1026-31
references.pmid: 9461463
references.citation: Lett Appl Microbiol. 2001 Mar;32(3):171-5
references.pmid: 11264747
references.citation: Nucleic Acids Res. 2000 Jun 15;28(12):E63
references.pmid: 10871386
references.citation: J Biochem Biophys Methods. 2007 Apr 10;70(3):499-501
references.pmid: 17011631
references.citation: FEMS Microbiol Lett. 2006 Jan;254(1):75-80
references.pmid: 16451182
references.citation: Clin Chem. 2001 Sep;47(9):1742-3
references.pmid: 11514425
references.citation: J Clin Microbiol. 2004 Nov;42(11):5349-52
references.pmid: 15528740
references.citation: Clin Microbiol Infect. 2007 Oct;13(10):980-4
references.pmid: 17803751
references.citation: FEMS Microbiol Lett. 2008 May;282(1):15-21
references.pmid: 18355290
references.citation: J Clin Microbiol. 2009 Jan;47(1):230-4
references.pmid: 19020065
references.citation: Pediatr Infect Dis J. 2008 Dec;27(12):1122-3
references.pmid: 18978513
references.citation: J Food Prot. 2005 Jan;68(1):146-9
references.pmid: 15690816
references.citation: J Clin Microbiol. 2005 Jun;43(6):2895-903
references.pmid: 15956414