PCRmax Pasteurella multocida Instruction Manual

Manganese-dependent
superoxide dismutase
(sodA)

Pasteurella multocida

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Pasteurella multocida is a small, Gram-negative, non-motile, non–spore-forming,
facultatively anaerobic coccobacillus. It often exists in the upper respiratory tract of
vertebrates including cats, dogs, cows, rabbits, pigs and fowl. It is the causative agent of
fowl cholera in chickens and turkeys, hemorrhagic septicemia in cattle and atrophic rhinitis
in pigs. The genome consists of a circular chromosome which is approximately 2250 kb
long, as well as a plasmid. The genome contains 129 lipo-proteins that are secreted and
located in the outer membrane. The bacterium has a capsule which helps it to avoid
phagocytosis, as well as lipopolysaccharides.

Pasteurella multocida can be transmitted through ingestion or inhalation, either during
direct contact or via fomites such as contaminated feed and water. Infection in humans is
often associated with an animal bite, scratch, or lick, but infection without epidemiologic
evidence of animal contact may occur. Biting arthropods do not seem to be important in
the epidemiology of this disease.

A major virulence factor in P. multocida is a bacteriophage encoded toxin. The toxin has
associated cell adhesion molecules to mediate attachment and receptor mediated
endocytosis into a number of host cell types, where it activates Rho GTPases. Rho
GTPases are associated with a number of host signalling pathways including those leading
to actin stress fiber formation, which subsequently aides endocytosis of the P. multocida
cells. The host cell cycle is also modulated by this toxin.

Pasteurella multocida does not remain viable for long periods in the environment, but it can
survive for hours and possibly days in damp soil or water. Rainy conditions and high
humidity facilitate transmission. The incubation period is usually 3 to 5 days but some
animals can carry the organism for varying periods without symptoms. Symptoms in
humans can include nasal discharge, sneezing, congestion and conjunctivitis.

Introduction to Pasteurella multocida

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MIN

MAX

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

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

• P.multocida specific primer/probe mix (150 reactions BROWN)

FAM labelled

• P.multocida 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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