PCRmax Candida albicans Instruction Manual

ribonuclease P RNA (RPR1)
gene

Candida albicans

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Candida albicans is a single-celled, diploid fungus and is part of the natural human gut
flora but as an opportunistic pathogen can also cause oral and genital infections.

C. albicans, which is present in around 80% of the population without causing infection,
can change from a unicellular organism to a pathogenic, multi-cellular, filamentous form. In
the commensal form, the fungus lives in the gastro-intestinal tract and growth is regulated
by other microorganisms as well as the host immune system. The change between these
forms, known as phenotypic switching may be regulated by differential gene expression
and regulation of certain transcription factors. Ultimately, the phenotypic switching to the
multi-cellular form allows penetration of the mucosal membrane by which infection occurs.

Infection by C. albicans causes Candidiasis, which can be either superficial or systemic,
causing life threatening disease. Superficial infections cause symptoms such as itching
and irritation which can usually be cleared by using anti-fungal drugs. However, in
immuno-suppressed individuals systemic infection occurs when the fungus penetrates the
mucosal membrane of the gut subsequently allowing release of toxins into the
bloodstream. Initially the infection presents with symptoms such as indigestion, nausea,
heartburn, constipation and/or diarrhea. As the infection progresses and the fungus enters
the blood stream, symptoms such as depression, lethargy, and muscle aches.

Introduction to Candida albicans

2

MIN

MAX

The C.albicans primers have been designed for the specific and exclusive in vitro
quantification of all C. albicans isolates and do not detect other Candida species. The
target sequence (RPR1) has previously been shown to be a good genetic marker for C.
albicans in other real time PCR based studies (Innings A 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 Candida albicans (C.albicans) genomes is designed for the in
vitro quantification of C.albicans genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the C.albicans genome.

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

3

Kit Contents

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

FAM labelled

• C.albicans 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

4