PCRmax Klebsiella pneumoniae Instruction Manual

outer membrane phosphate porin
precursor (phoE) gene

Klebsiella pneumoniae

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Klebsiella pneumoniae is a rod-shaped, non-motile, Gram-negative bacterium of the
Enterobacteriaceae family. The bacterium is around 2µm by 0.5µm in size which includes
a 160nm thick polysaccharide capsule known as antigen K which along with
lipopolysacchardies (antigen O) contributes to pathogenicity. The genome of this bacterium
is around 5Mbp in length and is arranged in a circular chromosome.

K. pneumoniae is found in the gastro-intestinal tract of mammals as part of the natural
flora. However, the bacterium can colonise the skin, pharynx and gastro-intestinal tract by
evading the host immune response. The polysaccharide capsule protects the bacterium
against phagocytosis by host polymophonuclear granulocytes and provides protection
against bactericidal host serum. The polysaccharide layer also contains adhesions that
mediate binding to the host cell. Individuals with impaired respiratory defenses are known
to be susceptible to infection of the lungs by this bacterium where necrosis, inflammation
and haemorrhaging of the lung tissue result in disease. K. pneumoniae is also known to be
a nosocomial infection which can lead to pneumonia, bacteremia and meningitis.

Introduction to Klebsiella pneumoniae

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MIN

MAX

The K.pneumoniae primers have been designed for the specific and exclusive in vitro
quantification of all K. pneumoniae isolates including the subsp. Rhinoscleromatis. The
primers and probe also detects Klebsiella variicola which is a potentially new and closely
related pathogenic Kelbsiella strain. The target sequence (phoE) has previously been
shown to be a good genetic marker for K. pneumoniae in other real time PCR based
studies (K.E. Shannon et.al 2007). 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 Klebsiella pneumoniae (K.pneumoniae) genomes is designed
for the in vitro quantification of K.pneumoniae genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the K.pneumoniae genome.

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

• K.pneumoniae specific primer/probe mix (150 reactions BROWN)

FAM labelled

• K.pneumoniae 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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