Use of Real-Time Polymerase Chain Reaction and Molecular Beacons for the Detection of Escherichia coli O157:H7

摘要: Abstract Molecular beacons (MBs) are oligonucleotide probes that fluoresce upon hybridization. In this paper, we described the development of a real-time PCR assay to detect presence Escherichia coli O157:H7 using these fluorogenic reporter molecules. MBs were designed recognize 26-bp region rfbE gene, coding for an enzyme necessary O-antigen biosynthesis. The specificity MB-based was evaluated various enterohemorrhagic (EHEC) and Shiga-like toxin-producing (STEC) E. strains as well bacteria species cross-react with O157 antisera. All serotype tested positively identified while all other species, including closely related O55 not detected by assay. Positive detection demonstrated when >10 2 CFU/ml present in samples. capability raw milk apple juice demonstrated. As few 1 after 6 h enrichment. These assays could be carried out entirely sealed tubes, enabling rapid semiautomated food environmental