PCRmax Mycoplasma haemofelis Instruction Manual

Heat shock protein 70 (dnaK)
gene

Mycoplasma
haemofelis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Mycoplasma haemofelis, previously known as Haemobartonella felis, is a Gram negative,
aerobic parasite responsible for several diseases in cats. They belong to the genus
Mycoplasma and class Mollicutes. Cats infected with M. haemofelis suffer from massive
bacteremia of red blood cells that leads to a severe haemolytic anaemia. The bacterium
lacks a cell wall and has a linear double stranded DNA genome of approximately 1.2 Mb in
length. Because of their small genome, the metabolic pathways are reduced, leading to a
heavy reliance on the nutrients afforded by the host environment.

M. haemofelis can be transmitted vertically from mother to offspring but is primarily
transmitted through bites from infected fleas and ticks. Once the parasite enters the blood
stream, it attaches to host red blood cells with polar filaments before invading the cells and
triggering the cat's immune response, releasing antibodies. The antibodies bind to the
foreign M. haemofelis proteins as a coating, which serves to mark the infected red blood
cell for removal and destruction. Due to the large numbers of infected red blood cells being
destroyed, the cat becomes anaemic and pale.

M. haemofelis is the causative agent for a range of feline diseases such as feline
conjunctivitis, lower respiratory tract infections, feline infectious peritonitis and feline
infectious anaemia (FIA). Clinical signs of the infection include anorexia, depression,
lethargy, weakness, fever and weight loss. Cats that have been infected with M.
haemofelis may remain permanent carriers and infection can re-emerge through stress.

Introduction to Mycoplasma haemofelis

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MIN

MAX

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

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

• M.haemofelis specific primer/probe mix (150 reactions BROWN)

FAM labelled

• M.haemofelis 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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