PCRmax Ehrlichia canis Instruction Manual

major outer membrane protein
(p30-10) gene

Ehrlichia canis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Ehrlichia canis is a small spherical intracellular bacterium with a single circular
chromosome. This bacterium acts as a causative agent of Ehrlichiosis, a disease
commonly affecting canine species. Canine ehrlichiosis is found worldwide but most
prominent in South America, Asia and Africa.

Ehrlichia are transmitted by ticks including the Brown Dog Tick, Rhipicephalus sanguineus
and the Lone Star Tick Amblyomma americanum. The primary vector of E. canis, the
brown dog tick, transfers the pathogen between hosts during blood meals. Ticks become
carriers of E. canis by feeding on an infected animal, if infected in the larval stage, the tick
can retain the bacteria in the midgut and salivary glands and infect hosts during blood
meals in both the nymph and adult stage of their life cycle. Due to the nature of this
pathogens cell walls it is able to resist the immune response of the host by evading the
hosts antibodies, E. canis penetrates the monocytes and macrophages of hosts and is
capable of spreading throughout the lymphatic system. The disease can take a chronic
form where bone marrow will fail to develop along with anaemia, leaving the animal
susceptible to further infection.

Dogs with Ehrlichiosis can be asymptotic, howevercommonly present with fever, lethargy,
depression, anorexia, and weight loss. Occasionally dogs may show symptoms of
neurologic disease, vomiting, or diarrhea. Humans can become infected with Ehrlichiosis,
but through a tick bite rather than through and infected dog. Common symptoms include
fever, chills, headache and muscle aches.

Introduction to Ehrlichia canis

2

MIN

MAX

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

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

3

Kit Contents

• E.canis specific primer/probe mix (150 reactions BROWN)

FAM labelled

• E.canis 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

4