PCRmax Mycoplasma gallisepticum Instruction Manual

cytadhesin protein (mgc2)

Mycoplasma
gallisepticum

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Mycoplasma Gallisepticum is a flask-shaped bacterium of the Mycoplasma genus.
Infections with this virus causes chronic respiratory disease in chickens and infectious
sinusitis in turkeys and other fowl. M. Gallisepticum lack a cell wall with the cells being
bound only by a plasma membrane. The double-stranded DNA genome is over 1M bp in
length arranged in a circular conformation and encoding over 800 genes. The largest
family of genes is the VlhA family which code pMGA lipoproteins involved in antigenic
variation.

M. Gallisepticum can survive in the environment for up to 2 months depending on the
conditions. This enables transmission via dust, drinking water and food as well as by direct
contact with an infected bird. The bacteria target the epithelial cells of the tracheal lumen,
adhering to the sialic acid residues with this cytadherence being mediated by two genes in
particular: CrmA and GapA. Once attached to the epithelial cells, the host cell membrane
fuses with the bacterium leading to infection with binary fission resulting in an increase in
Mycoplasma number within the cell within 24 hours of infection. Secondary infection with
other bacteria results from ciliostasis and deciliation of the tracheal epithelium which allows
access to erythrocytes.

Symptoms of M. Gallisepticum infection usually last for several weeks with resultant
mortality rates as high as 60% and those birds which do recover from infection remaining
carriers. Infection initially presents with a nasal discharge followed by a foamy or bubbly
exudate in the eyes as a result of swelling of nasal passages, sinuses, trachea, and
bronchi. The infection subsequently results in a persistent cough, sneezing and sniffing
with weight loss and a decrease in egg production also being apparent. Infected birds can
be treated with antibiotics such as Erythromycin, Tylosin, Spectinomycin or Lincomycin
administered via feed, water or injection.

Introduction to Mycoplasma gallisepticum

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MIN

MAX

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

The primers and probe sequences in this kit have 100% homology with a broad range of
M.gallisepticum 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.gallisepticum specific primer/probe mix (150 reactions BROWN)

FAM labelled

• M.gallisepticum 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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