PCRmax Vibrio cholerae (all subspecies) Instruction Manual

Outer membrane protein W
(ompW) gene

Vibrio cholerae all
subtypes

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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V. cholerae is a “comma” shaped Gram negative bacteria with a single polar flagella. There
are numerous strains of V. cholerae, some of which are pathogenic and some of which are
not. The complete genome consists of two chromosomes in a circular confirmation.
Chromosome 1 is 2.9 Mb long with 2,770 open reading frames (ORfs) and chromosome 2
is 1 Mb long with 1,115 ORFs. Pathogenic V. cholerae contains a genomic pathogenicity
island and is lysogenized with phage DNA, also adding to the pathogenicity of the
organism.

Transmission to humans occurs through the process of ingesting contaminated food or,
most commonly, water as it thrives in surface water. On ingestion the bacteria become
associated with the villi of the small intestine. There they produce cholera toxin, an
enterotoxin that leads to fluid secretion by the mucosal epithelial lining of the small
intestine, which 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 all subtypes

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MIN

MAX

Our kit for V.cholerae_subsp cluster has been designed for the specific and exclusive in
vitro quantification of all Vibrio cholerae subtypes including pathogenic and non­pathogenic strains. The primers and probe sequences in this kit have 100% homology
with a broad range of clinically relevant reference sequences based on a comprehensive
bioinformatics analysis.

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

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

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Kit Contents

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

FAM labelled

• V.cholerae_subsp 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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