PCRmax Theileria equi Instruction Manual

Equi merozoite antigen 1 (ema-1)
gene

Theileria equi

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Theileria equi also known as Babesia equi is an intraerythrocytic, protozoan parasite that
causes the disease equine piroplasmosis. Although the disease is endemic to sub-tropical
regions of the world, it has been reported in many countries and continents, thus making
this disease a cause of great concern in the global horse industry. This disease can also
affect donkeys, mules and zebras. It has a very small genome at 6.6 kb which is
monomeric linear DNA in structure. Trophozoites of T. equi appear as oval, round,
elliptical, or spindle-shaped structures of approximately 3µm in diameter. The merozoite
stage appears as two or four pyriform parasites together within the erythrocyte, each with a
length of only 1.5 µm on average. When four T. equi merozoites are present together in
one cell, they frequently form a characteristic "Maltese cross".

The parasite is transmitted by ixodid ticks of several genera including Boophilus,
Hyalomma, Dermacentor and Rhipicephalus. The parasite enters the host in trophozoite
("ring") form and invades the red blood cells by apicoplast action, where it multiplies
through binary fission. This increase in parasitic number results in rupture of the
erythrocytes causing anaemia and facilitating protozoan spread to other cells. The
merozoite form is ingested by ticks when feeding and settle to the mid gut where they
multiply. T. equi then migrates to the salivary glands, ready for injection into a new host.

Incubation for the parasite is 12 to 19 days. The severity of the disease depends on
whether the horse was previously exposed to the agent and on immunity status. The
clinical signs of this disease are not pathognomonic and may vary considerably between
horses. They may exhibit apathy, lethargy, fever, anaemia, icterus, weight loss and
oedema of distal limbs. The disease is common where the vector is present in high
numbers and infected horses can remain as carriers for long periods

Introduction to Theileria equi

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MIN

MAX

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

The primers and probe sequences in this kit have 100% homology with a broad range of
T.equi 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.equi specific primer/probe mix (150 reactions BROWN)

FAM labelled

• T.equi 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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