Detection of E. coli pathogenic genes (stx1 , stx2 and eae) in food products, animal feedstuff and environmental samples. Pre-enrichment according to ISO 16654:2001 (recommended). For use in real time instruments with FAM and ROX channels. AOAC certified
BACKGROUND: Escherichia coli, can cause enteric/diarrheagenic or extra-intestinal infections in humans. Enteric E. coli that cause disease can present different virulence determinants, corresponding to different pathotypes. These include the Shiga toxin-producing E. coli (STEC), comprising the enterohaemorrhagic E. coli (EHEC), and the enteropathogenic E. coli (EPEC). STEC are characterized by the production of Shiga-like toxins - Stx - corresponding to stx1 and stx2 genes (also know as verotoxins, encoded by vtx1 or vtx2 genes). EPEC was the first recognized pathogenic group, and presently, continues to be a leading cause of diarrhea among infants from developing countries worldwide. EPEC carry the eae (intimin-coding) gene and have the ability to cause attaching and effacing lesions in hosts, but do not produce Stx. EHEC was originally defined as a subset of STEC, that were associated with watery diarrhea, hemorrhagic colitis, hemolytic-uremic syndrome, and that in addition to the stx-encoding genes, usually carry the attaching and eae gene. These pathogenic organisms are mostly transmitted through undercooked minced beef, inadequately pasteurized milk, cold sandwiches, sprouts, and vegetables. The need for rapid, accurate, and sensitive methods for the detection of these E. coli strains is a major food safety issue. Since conventional microbiological methods for their detection and identification are time-consuming. Today, there are approved methods for PCR based detection of pathogenic genes of Escherichia coli unique DNA sequences, in particular using real time PCR and specific fluorescent probes.
PRODUCT DESCRIPTION: SUPREME REAL TIME DETECTION E. coli uses real-time PCR for the detection of pathogenic E. coli associated with the pathotypes EPEC, STEC and the subgroup EHEC associated with the combination of the virulence genes stx1 and/or stx2 and eae in a simple, reliable, and rapid procedure. The assay is based on 5’ nuclease real time PCR reactions to amplify a unique genomic sequence in the target. Our carefully designed primers and probe ensure highest sensitivity and specificity. The kit consists of an assay mix for the detection of the target as well as a positive control and an internal control (IC). To minimize PCR cross-contamination,the Kit contains dUTP and uracil-N glycosylase (UDG)
PRODUCT FEATURES:
-Amplification and detection: eae and stx1/stx2 genes of E. coli pathotypes EPEC, STEC and the subgroup EHEC
-Real-time PCR with rapid hot-start Taq DNA polymerase
-Internal Control to exclude false-negative results
-Optimized to handle PCR inhibitors, containing UDG enzyme
-Higher stability, enzymes in separate tubes
-PCR- platforms: runs on all established standard real-time PCR- platforms
-Harmonized thermal profiles to run with other SUPREME BIOPREMIER kits simultaneously
-Channel Target: FAM, Channel IC: ROX
-Available formats: Kits with 50 reactions for each target (EPEC and STEC) or 5 reactions for each target (EPEC and STEC)
-AOAC-RI VALIDATION: License number 081902