UC-TIB-EHEC/C.diff

For Research Use Only. Not for use in diagnostic procedures. Others UC-TIB-EHEC C.diff RUO UC-TIB-EHEC/C.diff PID00000818 10 280 590 001 10280590001 UC-TIB-EHEC/C.diff UC-TIB-EHEC/C.diff Reagents, kits 192 reactions Not Available false This product detects Enterohemorrhagic Escherichia coli (E.coli) (EHEC) Shiga toxin 1 and 2 (stx1/2) genes and the virulence factor eae (E. coli attaching and effacing gene) encoding intimin. Enteropathogenic E. coli (EPEC) also carry the eae gene but lack stx1 and stx2. In addition the Clostridioides difficile (C. difficile) toxin gene TcdB is detected enabling, the identification of potentially pathogenic C. difficile strains. en Bacterial DNA is analyzed by real-time PCR, which amplifies specific gene fragments that are detected in four detection channels. Channel 01 uses Coumarin-labeled probes to detect either a 107 bp fragment of the stx2 genes (stx2a/b/c/d/e/g/h/i/j/k/l) or a 96 bp fragment of stx2f. Channel 02 uses FAM-labeled probes to detect a 111 bp fragment of the eaeA gene. Channel 03 uses HEX-labeled probes to detect an 88 bp fragment of the stx1 genes (stx1a/c/e) or an 85 bp fragment of stx1d. Channel 04 uses LC640-labeled probes to detect a 110 bp fragment of the C. difficile tcdB gene. The (+) Ctrl comprises plasmids with a 676 bp long synthetic fragment of stx2 gene, a 395 bp long synthetic fragment of eaeA gene, a 663 bp long synthetic fragment of stx1 gene and a 232 bp long synthetic fragment of tcdB gene from C.difficile (Accession number: M59432 position 324 - 999 for stx2 gene, MK761163 position 614 - 1008 for eaeA gene, EU754738 position 84 - 747 for stx1 gene and X92982 position 7419 - 7650 for tcdB gene from C.difficile respectively). en The limit of detection (LoD) for UC-TIB-EHEC/C.diff was determined by analysis of serial dilutions of plasmid DNA in cobas® PCR media (200 μL sample volume). The study demonstrates that the UC-TIB-EHEC/C.diff detects at least 70 genome equivalent copies or less per mL for stx1 and stx2, E. coli eaeA and C. difficile toxin gene tcdB, determined by LOGIT analysis with a hit rate of 95%. en

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