PCRmax Enteropathogenic Escherichia coli Instruction Manual
Intimin (eae) gene
Enteropathogenic
Escherichia coli
PCRmax Ltd qPCR test
TM
150 tests
For general laboratory and research use only
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Escherichia coli (E. coli) is a Gram-negative, rod-shaped bacterium that is commonly found
in the lower intestine of warm-blooded organisms. Enteropathogenic Escherichia coli
(EPEC) is a pathogenic sub-type that causes diarrhea in not only humans but rabbits, cats,
dogs and horses. Most E.coli consist of a 4.6MB DNA circular genome which contains
between 4000-5500 genes. Several unique attaching and effacing virulence factors have
been found to aid in the pathogenicity of EPEC and the prescence or absence of certain
genes will help classify whether a serotype is EPEC or Enterotoxigenic E. coli (ETEC),
Enteroinvasive E. coli (EIEC) or Enterohemorrhagic E. coli (EHEC). The cells are about 2.0
microns (μm) long and 0.5 μm in diameter.
Transmission is most often caused by food or water contaminated with human or animal
feces. EPEC can cause either a watery or bloody diarrhea, the former associated with the
attachment to, and physical alteration of, the integrity of the intestine. Bloody diarrhea is
associated with attachment and an acute tissue-destructive process often due to the
presence of a toxin.
Along with diarrhea, other symptoms include abdominal cramps, vomiting and abdominal
pain. Symptoms typically appear 36 hours after infection but depending on the strain, they
can appear as early as 18 hours or as late as 72 hours after infection. The symptoms
typically clear after 3 days but in infants can last up to 2 weeks.
Introduction to Enteropathogenic Escherichia coli
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MIN
MAX
The PCR Max qPCR Kit for Enteropathogenic Escherichia coli (EPEC) genomes is
designed for the in vitro quantification of EPEC genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the EPEC genome.
The primers and probe sequences in this kit have 100% homology with a broad range of
EPEC sequences based on a comprehensive bioinformatics analysis.
If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to help@pcrmax.com and our bioinformatics team will
answer your question.
Specificity
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Kit Contents
• EPEC specific primer/probe mix (150 reactions BROWN)
FAM labelled
• EPEC positive control template (for Standard curve RED)
• Internal extraction control primer/probe mix (150 reactions BROWN)
VIC labelled as standard
• Internal extraction control DNA (150 reactions BLUE)
• Endogenous control primer/probe mix (150 reactions BROWN)
FAM labelled
• RNAse/DNAse free water (WHITE)
for resuspension of primer/probe mixes and internal extraction control DNA
• Template preparation buffer (YELLOW)
for resuspension of positive control template and standard curve preparation
Reagents and equipment to be supplied by the user
Real-Time PCR Instrument
DNA extraction kit
This kit designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.
Lyophilised 2x qPCR Mastermix
This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.
Pipettors and Tips
Vortex and centrifuge
Thin walled 1.5 ml PCR reaction tubes
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