title: Rapid and sensitive detection of Listeria monocytogenes by loop-mediated isothermal amplification.
abstract: Loop-mediated isothermal amplification (LAMP) was designed for detection of Listeria monocytogenes, which is an important food-borne kind of pathogenic bacteria causing human and animal disease. The primers set for the hlyA gene consist of six primers targeting eight regions on specific gene. The LAMP assay could be performed within 40 min at 65°C in a water bath. Amplification products were visualized by calcein and manganous ion and agarose gel electrophoresis. Sensitivity of the LAMP assay for detection of L. monocytogenes in pure cultures was 2.0 CFU per reaction. The LAMP assay was 100-fold higher sensitive than that of the conventional PCR assay. Taking this way, 60 chicken samples were investigated for L. monocytogenes. The accuracy of LAMP was shown to be 100% when compared to the "gold standard" culture-biotechnical, while the PCR assay failed to detect L. monocytogenes in two of the positive samples. It is shown that LAMP assay can be used as a sensitive, rapid, and simple detection tool for the detection of L. monocytogenes and will facilitate the surveillance for contamination of L. monocytogenes in food.
authors.affiliation: Poultry Institute, Chinese Academy of Agricultural Sciences, Jiangsu, China. tangmengjun1980@163.com
authors.forename: Meng-Jun
authors.initials: MJ
authors.lastname: Tang
authors.forename: Sheng
authors.initials: S
authors.lastname: Zhou
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Zhang
authors.forename: Jun-Hua
authors.initials: JH
authors.lastname: Pu
authors.forename: Qing-Lian
authors.initials: QL
authors.lastname: Ge
authors.forename: Xiu-Jun
authors.initials: XJ
authors.lastname: Tang
authors.forename: Yu-Shi
authors.initials: YS
authors.lastname: Gao
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics*; D002645:Chickens; D004269:DNA, Bacterial / Q000032:analysis; D005516:Food Microbiology; D006360:Heat-Shock Proteins / Q000235:genetics*; D006460:Hemolysin Proteins / Q000235:genetics*; D008089:Listeria monocytogenes / Q000235:genetics / Q000302:isolation & purification*; D008088:Listeriosis / Q000517:prevention & control; D008460:Meat / Q000382:microbiology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D001427:Bacterial Toxins; D004269:DNA, Bacterial; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; C038705:hlyA protein, Listeria monocytogenes