PCRmax Mammalian babesiosis Instruction Manual

18S ribosomal RNA (18S) gene

Mammalian babesiosis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Babesia species are parasitic protozoans of the Apicomplexa phylum that infect organisms
causing a haemolytic anaemia known as babesiosis.

These protozoa use ticks of the Ixodidae family as a vector. During tick feeding on an
invertebrate host, the trophozoite form is injected into the host blood system resulting in
invasion of the erythrocytes by apicoplast action. This group of organelles secretes
enzymes which erode the cellular membrane of the red blood cells facilitating entry. Once
within the cells, binary fission takes place resulting in the rupture of the cells allowing
protozoan spread through the blood stream and causing anaemia. The merozoite form is
subsequently ingested by the tick vector in a blood meal where it locates to the midgut and
multiplies. Migration to the salivary glands facilitates passing of the parasite to a new host.

Infection with a Babesia species results in Babesiosis, a disease which presents with
different symptoms depending on the causative agent. Infection with Babesia can result in
increased body temperature, loss of appetite, malaise, anaemia, severe weight loss and
enlargement of the spleen and liver. If the central nervous system is affected, respiratory or
circulatory failure may occur causing seizures and ataxia.

Oral atovaquone with oral azithromycin is the most commonly used treatment for infection
with B. divergens and B. microti. However, supportive therapy using intravenous fluid
replacement is also important and severe anaemia can be treated with whole blood
transfusions. In severe cases, blood transfusions have been performed just to lower the
parasitic load in the individual.

Introduction to Mammalian babesiosis

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MIN

MAX

Our kit for Babesia species genomes has been designed for the detection of the Babesia
species that cause babesiosis in mammalian species. The 18S ribosomal gene, is the
ideal target to achieve a broad based detection profile within this genus. The primers and
probe sequences in this kit have 100% homology with a broad range of clinically relevant
reference sequences based on a comprehensive bioinformatics analysis.

This kit will detect B.gibsoni, B.canis canis, B.canis vogeli, B.canis rossi, B.crassa, B.
major, B.hongkongnensis, B.caballia, B.ovata, B.occultans, B.microti, B. divergens, B.
capreoli, B.odocoilei, and B.bigemina.

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

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

• M.babesiosis specific primer/probe mix (150 reactions BROWN)

FAM labelled

• M.babesiosis 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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