PCRmax Trypanosoma equiperdum Instruction Manual

germline variant surface antigen
(VSG-78 BC-3 ) pseudogene

Trypanosoma
equiperdum

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Trypanosoma equiperdum is a protozoan parasite of the Trypanosoma genus that causes
dourine in equines. The disease can be found the world over, as it does not require insect
vectors that are influenced by climatic factors. The parasites mainly affect horses, donkeys
and mules. Dourine is endemic in parts of Africa, Asia, South Eastern Europe and the
Americas. T. equiperdum has a linear DNA genome that is in the form of chromosomes.

The disease is transmitted almost exclusively during breeding. Transmission from stallions
to mares is more common, but mares can also transmit the disease to stallions. The
parasite can be found in the vaginal secretions of infected mares, mucous exudates of the
penis, and sheath of stallions. Periodically, the parasites disappear from the genital tract
and the animal becomes non-infectious for weeks to months. Non-infectious periods are
more common in the later stages of the disease. The incubation period can last from a few
weeks to several years and the clinical signs often develop over weeks or months.

The symptoms of the disease are variable, though classically there are three stages.
During the first stage the genitalia become swollen and in mares there is a discharge from
the vagina, and patchy loss of pigment in the mucosa of the vulva or penis. Slight fever
and a loss of appetite may occur. The second stage manifests approximately 1 month later
with round urticarial eruptions and plaques on the neck, chest, flanks and rump which
disappear after a few days, but may reoccur. In the third stage is the onset of paralysis,
involving various muscles, and spreading to the hind legs, causing incoordination.
Complete paralysis of the four legs and death may finally occur, though the disease can
last for 1-2 years. This is the classical evolution of the disease but, often, far milder
symptoms are observed, particularly in in long-standing endemic regions, and not all
stages are always apparent. There also appear to be differences in virulence between
strains.

Introduction to Trypanosoma equiperdum

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MIN

MAX

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

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

• T.equiperdum specific primer/probe mix (150 reactions BROWN)

FAM labelled

• T.equiperdum 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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