1. Intended Use
Enterohemorrhagic E. Coli (EHEC) real time PCR kit is used for the detection of Enterohemorrhagic
E. Coli (EHEC) in stool or water samples by using real time PCR systems.
2. Product Description
O157,O26 and O111 is the major O antigens of EHEC, and O157: H7 is considered to be the major
EHEC strain which has been isolated. Nowadays, studies have found that other strains are related
with hemorrhagic colitis . The strain isolated in Germany ( outbreak of EHEC in May,2011) have
been identified as serotype O104.
The study shows that the major virulence factor of EHEC includes Stx (Shiga toxin ), LEE (locus of
enterocyte effacement) pathogenicity island, and 60MDa large plasmids. Stx, formerly called Vero
toxin or Shiga-like toxin, presents in the epithelial cells of blood vessels and it can cause system
response, resulting in HUS (heomlytic uremic syndrome); There are two types of Stx, named Stx1
and Stx2, and EHEC contains at least one of them. Stx1 and stx2 is also the main virulence genes of
Enterohemorrhagic E. Coli (EHEC) real time PCR kit contains a specific ready-to-use system for the
detection of shiga toxin gene stx1 and stx2 by polymerase chain reaction in the real-time PCR
system. The master contains reagents and enzymes for the specific amplification of the stx DNA.
Fluorescence is emitted and measured by the real time systems´ optical unit. The detection of
amplified EHEC DNA fragment is performed in fluorimeter channel FAM with the fluorescent
quencher BHQ1. DNA extraction buffer is available in the kit and excreta or water samples are used
for the extraction of the DNA. In addition, the kit contains a system to identify possible PCR
inhibition by measuring the HEX/VIC/JOE fluorescence of the internal control (IC). An external
positive control (1×107copies/ml) contained, allows the determination of the gene load. For further
information, please refer to section 9.3 Quantitation.