PCRmax Microsporum canis Instruction Manual

PQ-loop repeat protein gene

Microsporum canis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Microsporum canis is a zoophilic dermatophyte which is responsible for dermatophytosis in
dogs and cats. They cause superficial infections of the scalp (tinea capitis) in humans and
ringworm in cats and dogs. They belong to the family Arthrodermataceae and are most
commonly found in humid and warm climates. They have numerous multi-celled
macroconidia which are typically spindle-shaped with 5-15 cells, verrucose, thick-walled,
often having a terminal knob and 35-110 by 12-25 µm. In addition, they produce septate
hyphae and microconidia and the Microsporum canis genome is estimated at 23 Mb.

The fungus is transmitted from animals to humans when handling infected animals or by
contact with arthrospores contaminating the environment. Spores are very resistant and
can live up to two years infecting animals and humans. They will attach to the skin and
germinate producing hyphae, which will then grow in the dead, superficial layers of the
skin, hair or nails. They secrete a 31.5 kDa keratinolytic subtilisin-like protease as well as
three other subtilisin-like proteases (SUBs), SUB1, SUB2 and SUB3, which cause damage
to the skin and hair follicle. Keratinolytic protease also provides the fungus nutrients by
degrading keratin structures into easily absorbable metabolites.

Infection leads to a hypersensitive reaction of the skin. The skin becomes inflamed causing
the fungus to move away from the site to normal, uninfected skin. This creates
characteristic circular lesions with healing at the centre and inflammation at the edge.
Younger animals tend to be more susceptible to ringworm as their immune systems have
not been developed completely and their defence mechanisms are limited.

Introduction to Microsporum canis

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MIN

MAX

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

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

• M.canis specific primer/probe mix (150 reactions BROWN)

FAM labelled

• M.canis 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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