PCRmax Borrelia garinii Instruction Manual

16S-23S ribosomal RNA
intergenic spacer

Borrelia garinii

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Borrelia garinii is a Gram negative aerobic bacterium which is a causative agent of Lyme
disease. It is considered to be the most common tick-borne bacterial disease in the world.
They belong to the spirochete class, genus Borrelia and have a single stranded linear DNA
genome of approximately 950kb in length. The genome contains approximately 969 genes,
of which 931 are protein coding, and has several accompanying plasmids. It is a helical
bacterium with an inner and outer membrane as well as a flexible cell wall. The cell is
normally 0.1-0.5µm wide but due to its cylindrical shape can be 5-20 µm long. It is
structurally distinct as the flagella are located inside the periplasm between the inner and
outer cell membranes. These allow the cell to travel through the tissue of its infected host
or vector, causing it to be highly invasive.

Borrelia garinii is primarily transmitted by ticks of the genus Ixodes which serves as both a
vector and a reservoir for the virus. The bacteria are transmitted to avian and mammalian
reservoir hosts and to humans by Ixodes ricinus. It is also maintained in an enzootic cycle
in seabirds by I. uriae, a tick found at high latitudes in both the Northern and Southern
Hemispheres. The ticks usually acquire and transmit the spirochaete form of the bacteria
by feeding on an infected host. The bacteria subsequently reside in the lumen of the
tick’s digestive tract, multiplying whilst remaining there during maturation through the
nymph and adult life cycle stages. The bacteria then migrate to the salivary glands
attaching themselves to the immunosuppressive tick salivary proteins. As an adult, the tick
feeds on a new host resulting in infection of that host at around 24 hours after attachment.
The bacterial infection begins at the site of the tick bite but can spread if untreated.

Infection in the skin causes a “bull’s eye” rash around the bite site, sometimes
accompanied by erythema migraine. The rash can be treated with a course of antibiotics
but if left untreated can lead to a disseminated infection through the bloodstream resulting
in neurological problems such as facial palsy or meningitis. A third stage of this infection is
persistent and can be seen several months after the initial infection, causing chronic
symptoms of the heart, brain, nerves and joints, including arthritis.

Introduction to Borrelia garinii

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MIN

MAX

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

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

FAM labelled

• B.garinii 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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