PCRmax Cryptosporidium Instruction Manual

Oocyst wall protein (COWP)
gene

Cryptosporidium

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Cryptosporidium is a protozoan pathogen of the Apicomplexa phylum and causes a
diarrheal illness called cryptosporidiosis. Cryptosporidium does not utilise an insect vector
and is capable of completing its life cycle within a single host.

Cryptosporidiosis is typically an acute short-term infection but can become severe and
non-resolving in children and immunocompromised individuals such as AIDS patients. The
parasite is transmitted by environmentally hardy cysts (oocysts) that, once ingested, excyst
in the small intestine and result in an infection of intestinal epithelial tissue. Transmission
can occur via cysts excreted in faeces entering water supplies. Treatment of water by
filtration can remove these cysts.

After infection, symptoms may manifest within 2 days and last up to 2 weeks. These
include a low fever, stomach pains and watery diarrhea. Some individuals may be
asymptomatic and are, along with those with symptoms, contagious for several weeks.

A number of species of Cryptosporidium infect mammals. In humans, the main causes of
disease are C. parvum and C. hominis (previously C. parvum genotype 1). C. canis, C.
felis, C. meleagridis, and C. muris can also cause disease in humans. The genome of
Cryptosporidium parvum was sequenced in 2004 and was found to be unusual amongst
Eukaryotes in that the mitochondria seem not to contain DNA.

Introduction to Cryptosporidium

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MIN

MAX

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

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

• Crypto specific primer/probe mix (150 reactions BROWN)

FAM labelled

• Crypto 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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