PCRmax Burkholderia pseudomallei Instruction Manual

Hypothetical protein gene

Burkholderia
pseudomallei

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Burkholderia pseudomallei is a rod shaped Gram-negative, aerobic bacterium of the
Burkholderiaceae family. This species has a genome split over two chromosomes which
vary in size between strains but are approximately 4.1Mbp and 3.2Mbp respectively.
Infection with B. pseudomallei causes Melioidosis, also known as Whitmore’s disease, an
endemic disease in Southeast Asia and Northern Australia.

This species is commonly found in soil and can be transmitted in several ways: either by
direct contact with contaminated soil via skin abrasions; by inhalation of contaminated dust
or aerosols or by ingestion of contaminated water. Flagella enable motion and a secretary
system for adhesions and surface polysaccharides aid adhesion to the host cell
membrane. Once within the host cell, actin in the cytoskeleton is polymerised resulting in
cell fusion and multinucleated giant cells. A high level of adaption along with apparatus to
pump antimicrobial agents from the host cell is responsible for the pathogenicity of this
species. The production of endo- and exo-toxins presumably also has a role in
pathogenicity although this has not been characterised.

B. pseudomallei infection commonly has an incubation period of around two to four weeks
but an asymptomatic infection can remain dormant for many years by adaption to limited
nutrient or acidic environments. Typically, Melioidosis is characterised by acute or localised
infection presenting with fever in addition to localised pain at site of infection or abscesses
of the liver or spleen.

Introduction to Burkholderia pseudomallei

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MIN

MAX

Our kit for B. pseudomallei has been designed for the specific and exclusive in vitro
quantification of this species. The hypothetical protein gene selected as the target for this
kit is not present in other Burkholderia species. 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 Burkholderia pseudomallei (B.pseudomallei) genomes is
designed for the in vitro quantification of B.pseudomallei genomes. The kit is designed to
have the broadest detection profile possible whilst remaining specific to the B.
pseudomallei genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
B.pseudomallei 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.pseudomallei specific primer/probe mix (150 reactions BROWN)

FAM labelled

• B.pseudomallei 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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