PCRmax Tritrichomonas foetus Instruction Manual

Beta-tubulin 1 gene

Tritrichomonas foetus

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Tritrichomonas foetus is a single celled flagellated protozoan parasite that reproduces by
binary fission. The parasite is 5-25µm in length and is spindle shaped with an undulating
membrane seen over the entire length of the body caused by a recurrent flagella. The
organism has four flagella that are used to produce a jerky forward motion. This organism
parasitizes both the reproductive tract of naturally bred cattle and the digestive tract of
domestic cats.

In cattle, the protozoa infects the female reproductive tract when transmitted from the
foreskin of the bull where the parasite resides. Due to an increase in artificial insemination
procedures, cases of cattle infection are decreasing.
In domestic cats, the protozoa infect and colonise the large intestine as motile
trophozoites. The infection is most likely to be transmitted via the fecal-oral route or direct
contact as the infection is mainly seen in colonies of cats. There is little useful treatment for
infected cats as the organism is resistant to most traditional anti-protozoal drugs, and the
use of antibiotics may prolong shedding whilst alleviating the symptoms but not eradicating
the parasite, which can persist for the lifetime of the cat.

Infection in cattle may cause infertility and is thought to be linked to spontaneous abortions
in the first trimester of pregnancy. Infection in cats results in colitis and chronic diarrhea
with mild to severe inflammation of the colon and infiltration of lymphocytes and plasma
cells. These symptoms may persist for up to 2 years but on average can be overcome in
nine months. There is however evidence that after the symptoms have finished, shedding
may still occur for several months.

Introduction to Tritrichomonas foetus

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MIN

MAX

The primers have 100% homology with the two principal reference sequences with the
NCBI database for beta-tubulin (AY277787.1 and AY277787.1).

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

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

FAM labelled

• T.foetus 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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