PCRmax Lyme disease Instruction Manual

RecA gene

Lyme Disease

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Lyme disease is an infectious disease caused mainly by three species of bacteria that use
deer ticks as a vector. Borrelia burgdorferi, B. afzelii and B. garinii are all members of the
genus Borrelia, a group of Gram-negative, helical bacteria with flagella. The linear DNA
based genome ranges from 905K to 931K base pairs and has several accompanying
plasmids which vary between species.

A larval tick usually picks up the spirochaete form of the bacteria from feeding on an
infected host. The bacteria subsequently reside in the lumen of the ticks digestive tract,
multiplying whilst remaining there during maturation through the nymph and adult life cycle
stages. 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 the new host at around 24 hours after attachment. The bacterial
infection begins in the skin at the site of the tick bite but can spread if untreated.

Infection in the skin causes a bulls 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 spread causing a disseminated infection through the bloodstream causing
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 chromic
symptoms of the heart, brain, nerves and joints including arthritis.

Introduction to Lyme Disease

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MIN

MAX

The kit detects Borrelia burgdorferi, Borrelia garinii and Borrelia afzelii which are the
most significant causative agents of lyme Disease. Other Borrelia species will not be
detected with this kit. The RecA target gene has previously been shown to be a good
genetic marker for these three species in other clinical real time PCR based studies (D. S.
Saidac et,al 2009). 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 Lyme Disease (Lyme Disease) genomes is designed for the in
vitro quantification of Lyme Disease genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the Lyme Disease genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
Lyme Disease 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

• Lyme Disease specific primer/probe mix (150 reactions BROWN)

FAM labelled

• Lyme Disease 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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