BactoReal® Kit E. coli Typing is a series of different real-time PCR kits for detection of virulence-associated genes of E. coli (genes stx1, stx2, eae and ehxA). Depending on which kit and which combinations of kits are used, they allow the typing of enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC) and of Shiga toxin (verotoxin) producing E. coli (STEC).
Tests were developed and validated for the ABI PRISM® 7500 (Fast) instrument (Life Technologies),
LightCycler® 480 (Roche) and Mx3005P® (Agilent), but are also suitable for other real-time PCR instruments.
They allow the rapid and sensitive detection of DNA of virulence-associated genes of E. coli purified from stool or other sample material (e.g. with the QIAamp DNA Stool Mini Kit). Furthermore, isolated bacterial colonies can be picket from the plate, isolated e.g. with InstaGene™ Matrix (Bio-Rad) and analysed. An internal positive control system for detection in Cy5 channel (667 nm) excludes false-negative interpretation of results due to inhibition of real-time PCR (see 8. Interpretation of PCR-data).