A highly specific and sensitive loop-mediated isothermal amplification method for the detection of Escherichia coli O157:H7

作者：Ravan Hadi; Amandadi Mojdeh; Sanadgol Nima

来源：Microbial Pathogenesis, 2016, 91: 161-165.

DOI：10.1016/j.micpath.2015.12.011

• Loop-mediated isothermal amplification
• LAMP assay
• Escherichia coli O157:H7
• Z3276 gene

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 23276 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.

• 出版日期2016-2