PCRmax Enterocytozoon bieneusi Instruction Manual

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Enterocytozoon
bieneusi

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Enterocytozoon bieneusi, a fungus of the microsporida phylum is an obligate intracellular
parasite that infects intestinal epithelial cells. The linear DNA genome is estimated at 40
Mb, organized in 3 or 4 chromosomes with a total of 3,804 genes, 1,702 of these encoding
functional proteins. This species infects human and other mammals causing
microsporidiosis and many associated opportunistic infections which have had a serious
economic impact on the swine industry particularly.

Infective E. bieneusi spores are hardy and can survive in the environment for long periods
of time. The oval shape is created by two extracellular walls surrounding a membrane
containing the sporoplasm, the infectious material. Upon ingestion with contaminated food
stuffs or water, the extrusion apparatus comprising an anchoring disk, polar tubule, and
polaroplast extend from the spore piercing the host cell membrane. The sporoplasm is
then injected through this apparatus into the host cell, primarily the intestinal epithelium.
Within the host cell, proliferation by sporogony occurs in the cytoplasm and is followed by
maturation. The resultant new spores cause host cell death by disrupting the host cell
membrane leading to release into the intestine allowing for subsequent infection of
surrounding cells or excretion and transmission.

E. bieneusi causes microsporidiosis in humans, specifically in immune-compromised
individuals and has been linked with the wasting and chronic diarrhoea seen in individuals
with AIDS. Other symptoms include urinary tract infections and bowel perforation, cough
and laboured breathing due to infection of the lungs, and inflammation of the brain and
muscle tissue in cases with the fungi has spread to other parts of the body. Fumagillin and
Albendazole have been effective in treatment to a certain extent along with treatments
utilising inhibitors of chitin synthase enzymes.

Introduction to Enterocytozoon bieneusi

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MIN

MAX

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

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

• E.bieneusi specific primer/probe mix (150 reactions BROWN)

FAM labelled

• E.bieneusi 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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