PCRmax Theileria mutans Instruction Manual

32 kDa immunodominant
piroplasm antigen gene

Theileria mutans

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

East Coast fever (theileriosis) is a disease of cattle, sheep and goats caused by the
protozoan parasite Theileria parva. A form of East Coast fever called corridor disease (T.
mutans) is observed when the organism is transmitted from the African buffalo to cattle by
ticks including Rhipicephalus appendiculatus and Haemaphysalis spp., and causes a
benign bovine theileriasis but infection of cattle by t. mutans is called Corridor disease.

The Theileria mutans genome contains a complex series of overlapping perfect repeats
ranging between 99 and 20 bp in size. Experiments where the T. mutans genome was cut
frequently with restriction enzymes resulted in large numbers of fragments predominantly
below 50 kb which suggested a much higher GC content than the related T. parva. Our kit
targets the 32 kDa immunodominant piroplasm antigen gene of T. Mutans

Theilerial diseases are tick-borne diseases and Cattle are infected when they are bitten by
ticks that have fed on infected buffaloes. Therefore, the disease only occurs in cattle
grazing pastures on which buffaloes are or have recently been present. Even the presence
of a single buffalo for a short period may cause an outbreak of disease among cattle.

The disease does not normally spread between infected cattle but remains a major
constraint to introducing buffalos to cattle farming areas. It is the second most important
disease transmitted from buffalos to cattle after foot and mouth disease.

Symptoms include the following: Fever (shown by depression, listlessness, standing
apart), Enlargement of glands/lymph nodes, Decreased milk production, Loss of appetite,
watery discharge from eyes, pale mucous membranes of eyes/gums, weakness(difficulty
walking), cough, suppressed breathing, froth from nose/fluid in lungs and disorientation. 3

-4 days after symptoms present there is an 80% mortality rate.
Due to the rate of progression treatment is not usually possible, as it is not transmitted
between cattle, fastidious tick control and removal of animals from tick infested areas are
the best means of containment.

Turning sickness is an unusual form of theileriosis occurring when the parasites in infected
cells block blood vessels in the brain, leading to brain damage.(cerebral theileriosis)
resulting in confused animals which turn in circles (hence ‘turning sickness’) Turning
sickness it is serious and usually results in death but is less common. Redness and
bleeding of the brain/brain lining are present post mortem and can be diagnostic factors.

Introduction to Theileria mutans

2

MIN

MAX

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

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

3

Kit Contents

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

FAM labelled

• T.mutans 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

4