PCRmax Theileria annulata Instruction Manual

merozoite-piroplasm surface
antigen (Tams1) gene

Theileria annulata

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Theileria annulata is an obligate, intracellular protozoan of the Theileria genus. T. annulata
has an 8.3 megabase genome, comprising four haploid chromosomes. Infection with this
parasite causes tropical theileriosis or Mediterranean theileriosis in cattle, yak (Bos
gruniens) and water buffalo (Bubalus bubalis).

T annulata is transmitted by ticks of the Hyalomma genus. Protozoal sporozoites are
formed in the salivary glands of infected ticks and are subsequently transmitted to cattle
during feeding. Within 5-8 days of transmission Schizonts, also known as Koch‘s blue
body, form in the host lymph nodes. The schizonts number subsequently increases
enabling them to invade numerous erythrocytes where they become piroplasms.
Piroplasm-containing erythrocytes are ingested by uninfected larval ticks during feeding. T.
annulata undergoes a sexual cycle in the gut of the tick which allows it to reach the salivary
glands to complete the life cycle.

After an incubation period of 10 to 25 days clinical signs of infection develop including
Lymphoproliferative diseases, characterized by fever, leucopenia and anaemia, swelling of
the superficial lymph nodes, lethargy and diarrhoea. Infected animals may also develop
pulmonary edema, severe dyspnea, and a frothy nasal discharge. Some animals may
recover from theileriosis and become asymptomatic carriers while others may have poor
productivity and stunted growth. Animals with theileriosis are given acaricidal treatment.
They can also be immunized by inoculating them with live, attenuated parasites followed
by treatment with Terramycin LA.

Introduction to Theileria annulata

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MIN

MAX

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

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

FAM labelled

• T.annulata 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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