title: Development of double loop-mediated isothermal amplification to detect Listeria monocytogenes in food.
abstract: In this study, a double loop-mediated isothermal amplification (dLAMP) based on two target genes hlyA and iap was developed for the rapid detection of Listeria monocytogenes in food. The results revealed that the detection time and temperature of our dLAMP assay for L. monocytogenes were 15 min and 63 °C respectively, with a sensitivity of 10 fg DNA of L. monocytogenes per tube. While normal LAMP (nLAMP) of hlyA or iap was 100 fg DNA of L. monocytogenes per tube for 45 min and 63 °C. Furthermore, mineral oil and GoldViewII nucleic acid stain were chosen as the basic materials to develop a simple visualized identification of the positive samples. A total of 450 food samples were tested for L. monocytogenes using the dLAMP protocol developed in this study. The results showed that the accuracy of the dLAMP and the "gold standard" culture-biotechnical method were 100 % identical, suggesting that the modified dLAMP assay would provide a potential for detection of L. monocytogenes in food products.
authors.affiliation: Food Biotechnology and Food Safety Laboratory, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
authors.forename: Rina
authors.initials: R
authors.lastname: Wu
authors.forename: Xiang
authors.initials: X
authors.lastname: Liu
authors.forename: Bangcheng
authors.initials: B
authors.lastname: Guo
authors.forename: Fusheng
authors.initials: F
authors.lastname: Chen
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Wang
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D001431:Bacteriological Techniques / Q000379:methods*; D005516:Food Microbiology / Q000379:methods*; D006360:Heat-Shock Proteins / Q000235:genetics; D006460:Hemolysin Proteins / Q000235:genetics; D008074:Lipoproteins / Q000235:genetics; D008089:Listeria monocytogenes / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D008074:Lipoproteins; C116063:P60 protein, bacteria; C038705:hlyA protein, Listeria monocytogenes
references.citation: J Appl Microbiol. 2010 Apr;108(4):1380-8
references.pmid: 19796096
references.citation: J Clin Microbiol. 2008 Apr;46(4):1259-67
references.pmid: 18234868
references.citation: Int J Food Microbiol. 2006 Mar 15;107(2):202-6
references.pmid: 16289408
references.citation: Int J Food Microbiol. 2003 Jul 25;84(2):217-24
references.pmid: 12781944
references.citation: Microbiol Res. 2010 Mar 31;165(3):211-20
references.pmid: 19515543
references.citation: Food Microbiol. 2010 Feb;27(1):19-23
references.pmid: 19913686
references.citation: Int J Food Microbiol. 2011 Aug 2;148(2):75-9
references.pmid: 21652102
references.citation: Biochem Biophys Res Commun. 2001 Nov 23;289(1):150-4
references.pmid: 11708792
references.citation: J Clin Microbiol. 2004 Jan;42(1):257-63
references.pmid: 14715762
references.citation: Curr Microbiol. 2011 Dec;63(6):511-6
references.pmid: 21935669
references.citation: Int J Food Microbiol. 2010 Aug 15;142(1-2):60-6
references.pmid: 20584557
references.citation: J Biochem Biophys Methods. 2004 May 31;59(2):145-57
references.pmid: 15163526
references.citation: Parasit Vectors. 2011 Jun 21;4:115
references.pmid: 21693031
references.citation: World J Microbiol Biotechnol. 2012 Feb;28(2):523-31
references.pmid: 22806847
references.citation: Nucleic Acids Res. 2000 Jun 15;28(12):E63
references.pmid: 10871386
references.citation: J Microbiol Methods. 2012 Jun;89(3):179-84
references.pmid: 22475582
references.citation: Biochem Biophys Res Commun. 2002 Feb 1;290(4):1195-8
references.pmid: 11811989
references.citation: J Microbiol Methods. 2007 Dec;71(3):281-7
references.pmid: 18029039
references.citation: J Microbiol Methods. 2010 Feb;80(2):134-7
references.pmid: 19958798
references.citation: Int J Food Microbiol. 1996 Sep;32(1-2):73-90
references.pmid: 8880329
references.citation: Mol Biol Rep. 2012 Jan;39(1):445-9
references.pmid: 21573733
references.citation: Mol Cell Probes. 2010 Aug;24(4):190-5
references.pmid: 20230890