title: A highly specific and sensitive loop-mediated isothermal amplification method for the detection of Escherichia coli O157:H7.
abstract: E. coli O157:H7 is one of the most important foodborne pathogen that causes some human illnesses such as bloody diarrhea, hemolytic-uremic syndrome, and kidney failure. We developed a loop-mediated isothermal amplification (LAMP) assay with six special primers that target a highly specific 299-bp region of the Z3276 gene for the detection of E. coli O157:H7. Among 117 bacterial strains tested in this study, positive results were only obtained from E. coli O157:H7 strains. The sensitivity level of the Z3276-LAMP assay was determined to be 5 CFU/reaction tube in pure bacterial culture. Moreover, the LAMP assay was successfully applied to artificially contaminated ground beef with a sensitivity level of 10(3) CFU/mL without pre-enrichment and 10 CFU/mL after a 4-h pre-enrichment. In conclusion, the present LAMP assay would be a useful and powerful tool for the rapid, sensitive, and specific diagnosis of E. coli O157:H7 strains in resource limited laboratories.
authors.affiliation: Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran. Electronic address: ravan@uk.ac.ir.
authors.forename: Hadi
authors.initials: H
authors.lastname: Ravan
authors.affiliation: Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran.
authors.forename: Mojdeh
authors.initials: M
authors.lastname: Amandadi
authors.affiliation: Department of Biology, Faculty of Science, Zabol University, Zabol, Iran; Pharmaceutical Science Research Center (PSRC), Tehran University of Medical Sciences (TUMS), Tehran, Iran.
authors.forename: Nima
authors.initials: N
authors.lastname: Sanadgol
mesh_terms: D000818:Animals; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D004927:Escherichia coli Infections / Q000382:microbiology; D019453:Escherichia coli O157 / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis; D006801:Humans; D008460:Meat / Q000382:microbiology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers
keywords: Escherichia coli O157:H7; LAMP assay; Loop-mediated isothermal amplification; Z3276 gene