PCRmax Bacillus cereus E33 Instruction Manual

phage-related protein gene

Bacillus cereus E33

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Bacillus cereus is a Gram-positive, rod shaped, mesophilic bacterium of around 3-4um in
length which forms endospores. B.cereus has a circular chromosome with 5,411,809
nucleotides with over 5000 genes most of which are protein coding. This species also has
a range of plasmids that vary in size from 5K to 500K bp. B.cereus is the causative agents
of Rapid-onset emetic syndrome and Slow onset diarrhoeal syndrome, food poisoning
syndromes.

Transmission of B. cereus endospores is mainly via consumption of contaminated food
which has not been cooked properly. Ingestion results in spore germination and
subsequent infection of the small intestine where the bacteria produce 3 different
enterotoxins: hemolysin BL (HBL), non-hemolytic enterotoxin (Nhe) and Cytotoxin K (EntK)
which insert into the host cellular membrane forming pores via the beta-barrel
conformation. B. cereus also produces an emetic toxin (ETE) which has a ring shaped
structure and is heat resistant up to 120 degrees so is not easily destroyed during cooking.

Infection with this bacteria results in diarrhoeal syndrome presenting with symptoms
including watery diarrhoea, abdominal cramps and pain around 6 hours after initial
infection and lasting up to 24 hours. Alternatively emetic syndrome causes nausea and
vomiting with symptoms appearing much quicker but also lasting up to 24 hours. B. cerues
can also cause local and systemic infections especially in immunocompromised patients,
newborns and patients with surgical wounds.

Introduction to Bacillus cereus E33

2

MIN

MAX

Our kit for Bacillus cereus E33 has been designed for the specific and exclusive in vitro
quantification of the Bacillus cereus E33 strain alone. The target sequence, phage
related protein gene, is not known to be present in any other Bacillus species or B. cereus
strains.

The PCR Max qPCR Kit for Bacillus cereus E33 (B.cereusE33) genomes is designed for
the in vitro quantification of B.cereusE33 genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the B.cereusE33 genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
B.cereusE33 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

Kit Contents

• B.cereusE33 specific primer/probe mix (150 reactions BROWN)

FAM labelled

• B.cereusE33 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

4