PCRmax Bordetella bronchiseptica and B. parapertussis Instruction Manual

Putative major fimbrial structural
subunit (fimA) gene

Bordetella
Bronchiseptica/Bordet
ella Parapertussis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Bordetella parapertussis:
Bordetella parapertussis is a small Gram-negative bacterium of the genus Bordetella that
is adapted to colonise the mammalian respiratory tract. Two lineage's of B. parapertussis
have been described. The first infects humans and is responsible for a minority of cases of
the disease pertussis (also known as whooping cough). The second, ovine, lineage causes
chronic non-progressive pneumonia in sheep.

Bordetella bronchiseptica :
Also know as tracheobronchitis, canine cough in dogs, and feline bordetellosis in cats, is a
highly contagious respiratory disease in cats and dogs caused by the bacteria Bordetella
bronchiseptica.
Bordetella bronchiseptica has a circular chromosome consisting of approximately
5,338,400 base pairs. It has one or more medium to large-sized plasmids depending on
the strain. Of the circular chromosome, 68.07% of the composition are GC complements.
Approximately 5,007 are coding sequences with an average gene size of 978 base pairs.
There are 3 rRNA operons as well as 55 tRNA operons. Bordetella bronchiseptica is a
minute, gram-negative rod-shaped coccobacillus about .5-1 micrometers in diameter and 5
micrometers in length. It may or may not have a flagella, dependent on environment.
Optimal growth temperature is 35-37°C.
Although it is known to colonize in respiratory tracts of animals, it can also withstand
surviving long-term in the environment. Bordetella cells have a 5-layered cell wall. The first
3 layers give the organism a lobulated appearance.

Bordetella spreads through direct contact (licking, nuzzling), through the air (coughing or
sneezing), or via contaminated fomites. Transmission can occur cross-species between
dogs and cats.

Introduction to Bordetella Bronchiseptica/Bordetella
Parapertussis

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MIN

MAX

The PCR Max qPCR Kit for Bordetella Bronchiseptica/Bordetella Parapertussis (B.
bronchiseptica/B.parapertussis) genomes is designed for the in vitro quantification of B.
bronchiseptica/B.parapertussis genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the B.bronchiseptica/B.parapertussis
genome.

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

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

FAM labelled

• B.bronchiseptica/B.parapertussis 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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