PCRmax Coxiella burnetii Instruction Manual

DNA gyrase subunit A

Coxiella burnetii

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Coxiella Burnetii is a Gram-negative non-sporulating, rod-like intracellular bacterium that is
responsible for human Q fever. The pathogen can be considered as a potential agent for
bioterrorism as it is highly infectious and resistant to heat and drying. The genome is
approximately 2 Mb long and is circular in conformation. C. burnetii possesses a
lipopolysaccharide structure that is important in virulence and responsible for antigenic
phase variation, analogous to the smooth-rough LPS variation seen in enteric Gram­negative microorganisms. Bacterial isolates from eukaryotic hosts have a phase I (smooth)
LPS, which can protect the microorganism from microbicidal activities of the host.

Coxiella burnetii can induce abortion in domestic mammals (cat, dog, rabbit) and
ruminants, and these animals represent their main reservoir. The danger posed by Coxiella
is that they can be excreted by animals exhibiting no apparent clinical signs of the disease,
whether following natural infection or because the animals have been vaccinated with an
ineffective vaccine. This can then be transmitted to humans via inhalation or tick bites.
After initial infection at the site of entry, the microorganism is engulfed by resident
macrophages and transported systemically, causing histopathological changes in the
lungs, liver, and spleen. After uptake by host cells, the acidic conditions within the
phagolysosome allow the bacteria to grow. Eventually, proliferation within the
phagolysosome leads to rupture of the host cell and infection of a new population of host
cells.

The incubation period can vary from a few days to several weeks and the severity of
infection varies in direct proportion to the infectious dose in normal individuals. There are
no characteristic symptoms of Q fever, but fever, severe headache, and chills tend to be
prevalent. Fever usually peaks at 40°C and lasts for approximately 13 days. Fatigue and
sweats also frequently occur. Pneumonia is a common clinical presentation. Cough,
nausea, vomiting, myalgia, arthralgia, chest pain, hepatitis, and occasionally,
splenomegaly, osteomyelitis, and meningoencephalitis are symptoms that are also
associated with acute Q fever. Fatalities in cases of acute Q fever are rare, with fewer than
1% of cases resulting in death. Children develop symptomatic disease less frequently than
adults.

Introduction to Coxiella burnetii

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MIN

MAX

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

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

• C.burnetii specific primer/probe mix (150 reactions BROWN)

FAM labelled

• C.burnetii 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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