PCRmax Vibrio (all species) Instruction Manual

RNA polymerase alpha subunit
(rpoA) gene

Vibrio all species

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Vibrio is a genus of “comma” shaped, facultatively anaerobic Gram negative bacteria that
are commonly present in marine environments. These bacteria are motile by means of a
single polar flagellum, they do not form spores and can undergo both respiratory and
fermentative metabolism. They are sensitive to acidic pH but can tolerate alkaline pH very
well. Most species of Vibrio are halophilic and require salt water for growth. In tropical and
temperate regions, disease-causing species of Vibrio occur naturally in marine and coastal
environments. They are heterotrophic organism and derive their nutrients through a
mutualistic or parasitic relationship with the host organism.

The genus contains at least twelve species that are pathogenic to humans, eight of which
are associated with food-borne disease. The majority of food-borne disease is caused by
V. cholerae, V. parahaemolyticus and V. vulnificus. Some species such as V. cholerae and
V. parahaemolyticus are primarily associated with gastrointestinal disease, while others
like V. vulnificus can cause non-intestinal illness, such as septicaemia. While
immunosuppressed individuals are most susceptible to Vibrio infections, these bacteria are
capable of being harmful to healthy individuals. Pathogenic Vibrios, in particular V.
cholerae, can also be recovered from freshwater reaches of estuaries, where it can be
introduced by faecal contamination as well as having a natural presence in this
environment.

Introduction to Vibrio all species

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MIN

MAX

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

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

• Vibrio_spp specific primer/probe mix (150 reactions BROWN)

FAM labelled

• Vibrio_spp 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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