PCRmax Vibrio cholerae (toxigenic subtypes) Instruction Manual

ctxB (cholera toxin) gene

Vibrio cholerae
toxogenic subtypes

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Cholera, sometimes known as epidemic cholera, is an infectious gastroenteritis caused by
the bacterium Vibrio cholerae. Transmission to humans occurs through the process of
ingesting contaminated water or food. The major reservoir for cholera was long assumed
to be humans themselves, but considerable evidence exists that aquatic environments can
serve as reservoirs of the bacteria.

V. cholerae is a Gram-negative bacterium that produces cholera toxin, an enterotoxin,
whose action on the mucosal epithelium lining of the small intestine is responsible for the
characteristic massive diarrhoea of the disease. In its most severe forms, cholera is one of
the most rapidly fatal illnesses known, and a healthy person may become hypotensive
within an hour of the onset of symptoms; infected patients may die within three hours if
treatment is not provided. In a common scenario, the disease progresses from the first
liquid stool to shock in 4 to 12 hours, with death following in 18 hours to several days
without rehydration treatment.

Introduction to Vibrio cholerae toxogenic subtypes

2

MIN

MAX

Our kit for V.cholerae has been designed for the specific and exclusive in vitro
quantification of strains responsible for causing cholera in humans.

Accession numbers for detected sequences
EF591295.1, CP000626.1, AE003853.1, EF158842.1, AY349176.1, X76390.1

The PCR Max qPCR Kit for Vibrio cholerae toxogenic subtypes (V.cholerae) genomes is
designed for the in vitro quantification of V.cholerae genomes. The kit is designed to have
the broadest detection profile possible whilst remaining specific to the V.cholerae genome.

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

• V.cholerae specific primer/probe mix (150 reactions BROWN)

FAM labelled

• V.cholerae 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