PCRmax Ehrlichia (all species) Instruction Manual

16S ribosomal RNA (18S) gene

Ehrlichia genus

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Ehrlichia is a genus of small, Gram-negative, obligatory intracellular bacteria belonging to
the Anaplasmataceae family. Ehrlichia genomes consist of a circular DNA chromosome of
about 1.1 to 1.7Mb varying between species. Ehrlichia infection targets blood cells causing
either Human granulocytic ehrlichiosis (HGE) or Human monocytic ehrlichiosis (HME)
based on the type of blood cell infected.
Ehrlichiosis use ticks as vectors for infecting mammals with lone star tick-associated
bacteria infecting monocytes and deer tick-associated bacteria infecting granulocytes. The
bacteria attach to the host cell via an interaction between the bacterial 120-kD surface­exposed glycoprotein and the host-cell expressed E and L selectins, triggering receptor­mediated endocytosis. The bacteria survives in the leukocyte phagosome (called morula),
where they divide by binary fission to produce microcolonies. The infected cell is destroyed
by the morula, releasing bacteria to infect nearby cells.
Symptoms of HME and HGE include sudden fever, head and muscle aches and in some
cases, a rash. In immunocompromised hosts, ehrlichiosis may be severe, manifesting as a
Rocky Mountain spotted fever (RMSF)–like illness that may be fatal. Prevention is mainly
avoiding exposure to ticks as there is no vaccine but Doxycycline is used in treatment after
infection.
The genus Ehrlichia comprises the species E. canis, E. chaffeensis, E. ewingii, E. muris
and E. ruminantium.

Introduction to Ehrlichia genus

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MIN

MAX

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

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

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

FAM labelled

• Ehrlichia_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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