PCRmax Rickettsia (all species) Instruction Manual

Citrate synthase (gltA) gene

Rickettsia (all species)

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Rickettsia is a genus of Gram-negative bacteria which are pleomorphic, motile, rod-shaped
and obligate intracellular parasites. The genomes of Rickettsia species are a single circular
DNA chromosome typically up to 1Mbp. These genomes consist of 900 to 1500 genes
depending on the species, of which 704 protein coding and 39 RNA-coding genes are
common to all Rickettsia genomes sequenced to date. Rickettsia species can be classified
into the following groups based on the severity of the infections they cause: Spotted Fever
group; Typhus group and Scrub Typhus group (Tsutsugamushi disease).
Rickettsia species are carried by many ticks, fleas and lice and are transferred to humans
via bites from infected arthropods. These bacteria adhere to the endothelial lining of the
vasculature within the various organs affected utilising adhesins thought to be outer
membrane proteins, subsequent phagocytosis results in host cell invasion. Once inside the
cell the bacteria can multiply resulting in lysis of the host cell due to accumulation of large
numbers of bacteria (typhus) or escape from the cells causing damage to the membrane
resulting in influx of water (spotted fever). Specific invasion of cells lining blood vessels
results in Rocky Mountain spotted fever (RMSF).
Infection with Rickettsia species can cause typhus, rickettsialpox, Boutonneuse fever,
Rocky Mountain spotted fever and Q fever with common symptoms including skin rash,
headache, altered conscious state, heart failure, and shock. RMSF specifically causes
haemorrhage, inflammation and blood clots. Infection with the most virulent Rickettsiae
species (R. rickettsii and R. Prowazekii) results in a high mortality rate if treatment is not
administered early in the infection. Tetracyclines are the drug of choice in treating most
Rickettsial infections. Arthropod and rodent control are possible to prevent infections but
often difficult. No vaccines are currently available.

Introduction to Rickettsia (all species)

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MIN

MAX

Our kit for Rickettsia has been designed for the specific and exclusive in vitro
quantification of rickettsia species within this genus. A highly conserved sequence within
the gtfA gene has previously been shown to be a good target sequence in other clinical
real time PCR based studies (J. Stenos et.al., 2005). 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 Rickettsia (all species) (Rickettsia_spp) genomes is designed
for the in vitro quantification of Rickettsia_spp genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the Rickettsia_spp genome.

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

• Rickettsia_spp specific primer/probe mix (150 reactions BROWN)

FAM labelled

• Rickettsia_spp 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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