PCRmax Mycoplasma hyopneumoniae Instruction Manual

RNA polymerase beta subunit
(rpoB) gene

Mycoplasma
hyopneumoniae

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Mycoplasma hyopneumoniae (M. hyopneumoniae) is a species of bacteria known to cause
Porcine Enzootic Pneumonia in pigs. It is a gram positive member of the Mycoplasma
genus. The cells are flask shaped and are between 400 and 1200nm in size. M.
hyopneumoniae genome is about 892 kbp long with G+C content of 28.6 mol%.

M. hyopneumoniae has been found to attach to the cilia of epithelial cells in the lungs of
swine. They cause cilia to stop beating (ciliostasis), clumping and loss of cilia, eventually
leading to epithelial cell death; which is the source of the lesions found in the lungs of pigs
with porcine enzootic pneumonia.
Direct contact with infected pigs has been established as the chief route of transmission,
constituting the main point of entry of the agent into the herd. Latently infected animals,
aerosol spread, and fomites are alternative routes of infection of naïve swine herds.
Livestock handlers may act as mechanical vectors in the transmission of the pathogen as
well.

Age has no influence on the expression of the clinical signs. In infected pigs, a slight
hyperthermia (40°C to 40.3°C) may be noticed one week after infection. Coughing begins
two weeks after infection, peaks after five weeks, then gradually declines. Twelve weeks
post infection, no clinical symptoms are noticeable. This bacterium is a concern in the
livestock industry as it causes a significant reduction in the growing weight of pigs.
Pneumonic lesions are formed within the lungs of infected animals as well.

Introduction to Mycoplasma hyopneumoniae

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MIN

MAX

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

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

FAM labelled

• M.hyopneumoniae 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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