PCRmax Escherichia coli (tellurite resistant) Instruction Manual

E.coli generic detection of beta­D-glucuronidase (uidA) gene
&
tellurite resistance (terC) gene

tellurite resistant
Escherichia coli

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Quantification of tellurite resistant Escherichia coli genomes.

Advanced kit handbook HB10.07.08

Published Date: 26/04/2016

Escherichia coli are one of many species of bacteria living in the lower intestines of
mammals, known as gut flora. When located in the large intestine, it assists with waste
processing, vitamin K production, and food absorption. Discovered in 1885 by Theodor
Escherich, a German pediatrician and bacteriologist, E. coli are abundant: the number of
individual E. coli bacteria in the faeces that a human defecates in one day averages
between 100 billion and 10 trillion. However, the bacteria are not confined to the
environment, and specimens have also been located, for example, on the edge of hot
springs. The bacteria are Gram-negative, rod-shaped, flagellated and non-spore forming.
Most strains are non-pathogenic but some cause food poisoning in humans with
transmission largely being through the faecal-oral route. E.coli have a circular, DNA
genome of approximately 4.6 Mb but also carry plasmids.

Tellurium is occasionally found native, but is more often found as the tellurite of gold, and
combined with other metals. Tellurite is toxic to most micro-organisms, especially Gram­negative bacteria. Tellurium compounds are used in the film and rubber industries and in
the manufacture of batteries, and are found in fairly large amounts in the human body.
Resistance to tellurite is usually mediated by conjugative plasmids and the determinants
encoded on these plasmids are usually highly specific for tellurite necessary for
resistance. Potassium tellurite (K2TeO3) is metabolized to form intracellular crystals of
black metallic tellurium, which are often deposited just inside the inner membrane.
Resistant cells grow as black colonies on relatively high concentrations of K2TeO3.
Tellurite resistance is caused by TerB, -C, -D, and -E genes and they are located on the O
island of inserted DNA segments.

The enteric E. coli (EC) are divided on the basis of virulence properties into
enterotoxigenic (ETEC – causative agent of diarrhea in humans, pigs, sheep, goats, cattle,
dogs, and horses), enteropathogenic (EPEC – causative agent of diarrhea in humans,
rabbits, dogs, cats and horses); enteroinvasive (EIEC – found only in humans),
verotoxigenic (VTEC – found in pigs, cattle, dogs and cats); enterohaemorrhagic (EHEC –
found in humans, cattle, and goats, attacking porcine strains that colonize the gut in a
manner similar to human EPEC strains) and enteroaggregative E. coli (EAggEC – found
only in humans).

Introduction to tellurite resistant Escherichia coli

2

Quantification of tellurite resistant Escherichia coli genomes.

Advanced kit handbook HB10.07.08

Published Date: 26/04/2016

MIN

MAX

The PCRmax qPCR Kit for tellurite resistant Escherichia coli (E.coli_TeR) genomes is
designed for the in vitro quantification of E.coli_TeR genomes. The kit is designed to have
the broadest detection profile possible whilst remaining specific to the E.coli_TeR genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
E.coli_TeR 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 [e-mail] and our bioinformatics team will
answer your question.

Specificity

3

Quantification of tellurite resistant Escherichia coli genomes.

Advanced kit handbook HB10.07.08

Published Date: 26/04/2016

Kit Contents

• beta-D-glucuronidase (uidA) gene primer/probe mix (150 reactions BROWN)

FAM labelled

• tellurite resistance (terC) gene primer/probe mix (150 reactions BROWN)

FAM labelled

• beta-D-glucuronidase (uidA) gene positive control template (for Standard curve
RED)

• tellurite resistance (terC) gene 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 templates and standard curve preparation

Reagents and equipment to be supplied by the user

Real-Time PCR Instrument

DNA extraction kit

This kit is 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

4

Quantification of tellurite resistant Escherichia coli genomes.

Advanced kit handbook HB10.07.08

Published Date: 26/04/2016