EHEC/EPEC PCR Kit

The EHEC/EPEC PCR Kit has been tested on the Roche LightCycler 480II/480 z, Agilent Technologies Mx3005P, Applied Biosystems ABI 7500, Bio-Rad CFX96, and QIAGEN Rotor-Gene Q real-time PCR instruments. Contact ALPCO for addition real-time PCR instrument information. The EHEC/EPEC PCR kit is a multiplex real-time PCR assay for the direct, qualitative detection and differentiation of genes encoding the virulence-factors of EHEC, STEC, EPEC and EIEC/Shigella spp. After DNA isolation, amplification of the gene fragments specific for the virulence factors stx1/stx2, eae and ipaH (if present) occurs. The amplified targets are detected with hydrolysis probes, which are labelled at one end with a quencher and at the other end with a fluorescent reporter dye (fluorophore). In the presence of a target the probes hybridize to the amplicons. During the extension step the Taq-Polymerase breaks the reporter-quencher proximity. The reporter emits a fluorescent signal which is detected by the optical unit of a realtime PCR instrument. The fluorescence signal increases with the number of formed amplicons. The EHEC/EPEC assay contains an Internal Control DNA (ICD) as an internal control of sample preparation procedure and to determine possible PCR-inhibition.