PCRmax Babesia divergens Instruction Manual

18S ribosomal RNA gene

Babesia divergens

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Targeting the Internal transcribed spacer 1 gene

Babesia divergens is an intraerythrocytic protozoan parasite, transmitted by the tick Ixodes
ricinus. It is the main agent of bovine babesiosis, or "redwater fever", in Europe. Younger
cattle are less susceptible. Many species of Babesia only infect non-human mammalian
hosts, most commonly cattle, horses, and sheep. B. microti and B. divergens are the two
main pathogenic species in humans.
The current emphasis in Europe on sustainable agriculture and extensification is likely to
lead to an increase in vector tick populations with increased risk of infection.

The targeted region is the 18S ribosomal RNA gene

Babesia is spread through the saliva of a tick when it bites. At its nymph stage, a tick will
bite into the skin for a blood meal. The tick, if not removed, will stay attached for 3 to 6
days with longer periods of feeding associated with a higher probability of acquiring the
parasite. The parasite can survive in the tick as it molts through its various developmental
stages resulting in all stages being potentially infectious. Some species of Babesia can be
transmitted from a female tick to its offspring before migrating to salivary glands for
feeding.

Babesia is transmitted by inoculation after the bite of Ixodidae ticks. There are many
species of Ixodidae that transmit the disease. In the Americas Ixodidae scapularis is the
most common vector. As a protozoan parasite, an effective way to identify Babesia
infection though blood sample testing using qPCR. Historically, Babesiosis diagnosis was
carried out with xenodiagnosis in hamsters for B. microti and in gerbils for B.divergens.
While successful at identifying the disease, this diagnostic technique has been abandoned
for faster diagnostic measures.

Introduction to Babesia divergens

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MIN

MAX

The 18S ribosomal gene, is the ideal target to achieve a broad based detection profile for
all strains within this species. 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.

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

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

• B.divergens specific primer/probe mix (150 reactions BROWN)

FAM labelled

• B.divergens 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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