PCRmax Mycoplasma suis Instruction Manual

16S ribosomal RNA gene

Mycoplasma suis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Mycoplasma suis (previously known asEperythrozoonosis suis), is a sporadic disease,
usually observed in young, growing pigs, characterized by listlessness, fever, anorexia,
haemolytic anaemia and, in severe cases, by icterus (jaundice).

The disease is caused byM. suisan obligate, intracellular bacteria. The organism is
usually from 0.8 to 1.0 um in diameter but may appear larger during the later stages of
infection. It is predominantly spherical (coccid-shaped) but may appear rod or ring-shaped
when viewed on a cell membrane.

In utero transmission has been demonstrated experimentally and although M. suisoccurs
in all ages of swine, it is most visible in young pigs. Prevalence of less than 15% is
common within an outbreak and it appears that stressed animals are more susceptible.

M. suis is often concurrent with other diseases and more prevalent during warmer weather.
Since the disease occurs more frequently in the summer, it is suspected that mosquitoes
and biting flies, as well as the hog louse (Haematopinus suis), play a role in transmission.
The use of contaminated surgical instruments (including castration knives) and needles
used repeatedly during vaccination are believed to transmit the organism. Ongoing
infectious and parasitic diseases are believed to play an important role in precipitating
outbreaks

AlthoughM. suisrarely causes spectacular death losses, it is recognized as an occasional
cause of anaemia or icterus in young pigs.

OnceM. suisis introduced into the blood, a fever occurs in about 7-8 days. Infected
livestock are likely to become hypoglycaemic, which may lead to coma and death.
Haemolytic anaemia develops gradually, often requiring a week after onset before it is
clinically apparent. In outdoor conditions many affected animals recover but remain
carriers.
M. suisis an unlikely cause of major disease by itself.

Most t serologic tests have limited availability Real-Time PCR is highly sensitive and quick
to perform as well as being cost effective for mass screening.
Affected pigs can be treated individually with injectable broad spectrum antibiotics (e.g.
oxytetracycline) There are no vaccines for the prevention ofM. suis.

Introduction to Mycoplasma suis

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MIN

MAX

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

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

FAM labelled

• M.suis 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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