PCRmax Pneumocystis jirovecii Instruction Manual

Kexin-like serine protease (kex1)
gene

Pneumocystis jirovecii

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Pneumocystis jirovecii is a fungi of the Pneumocystidaceae family, the genome of which
has not been fully sequenced. This species of fungus causes Pneumocystis Pneumonia in
human hosts.

Inhalation of P. jirovecii allows entry to the host lungs where it adheres to the epithelium of
the alveoli. Here the cysts initially undergo meiosis and subsequently mitosis in three
stages allowing an increase of the intracystic nuclei from 2 to 8. At the end of the third
stage, excystment allows release of the trophic form of the protozoa which can be up to
4µm in diameter. The trophic form undergoes binary fission to produce many haploid
trophic forms that conjugate and produce early cysts.

Within the host, the protozoa only cause disease in immuno-supressed or immuno­compromised individuals. In these cases a form of pneumonia called Pneumocystis
Pneumonia (PCP) can be seen. This disease presents with symptoms including a cough,
shortness of breath and fever, although the infection can spread to other organs including
the liver, kidneys and spleen. A combination of antibiotics is typically used to treat the
infection with a treatment course running for up to 3 weeks.

Introduction to Pneumocystis jirovecii

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MIN

MAX

Our kit for Pneumocystis. Jirovecii cluster has been designed for the specific and
exclusive in vitro quantification of the Pneumocystis jirovecii species within human
samples. Please note that this species has recently been reclassified and was
previously known as P.carinii f. sp. Hominis. The target sequence within the KEX1 gene
has previously been shown to be a good genetic marker for P. jirovecii in other clinical
real-time PCR based studies (Rohner P, et.al 2009). This gene has no known homology
with other species in the P. carinii at the primer binding sites. 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 Pneumocystis jirovecii (P.jirovecii) genomes is designed for
the in vitro quantification of P.jirovecii genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the P.jirovecii genome.

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

FAM labelled

• P.jirovecii 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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