PCRmax Proteus mirabilis Instruction Manual

RNA polymerase beta subunit
(rpoB) gene

Proteus mirabilis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Proteus mirabilis is a Gram-negative, facultatively anaerobic, rod shaped bacterium. It can
be found as part of the micro flora in the human intestine. This organism is not usually a
pathogen, but does become a problem when it comes into contact with urea in the urinary
tract. From there, infection can spread to other parts of the body. Proteus can display two
different morphological and physiological forms; one is known as the swimmer cells and
the other as swarmer cells. In aqueous suspension Proteus mirabilis is found in the
swimmer state, which is a small rod-like cells1 to 2 μm in length. They contain 8 to 10
flagella that aid in their swimming motility. On contact with a surface, Proteus mirabilis
changes to the swarmer state where the cell considerably increases in length to form
highly flagellated filaments that are 20 to 80 μm in length. These cells line up in parallel to
form rafts that are able to move rapidly over surfaces en masse. On semi-solid surfaces
such as an agar surface, they form concentric rings of growth. This pattern is caused by
the coordinated burst of swarming activity interspeded with a consolidation to the swimmer
state. The genome is 4.06Mb that includes a pathogenicity island that assists in infection.
The bacterium also carries a plasmid that is approximately 36,000bp long.

Proteus mirabilis is a significant pathogen of the urinary tract. Urinary tract infection is a
process following certain steps; it first starts with colonization of the bladder, which causes
cystitis. Then, the infection proceeds to the kidneys, which leads to acute pyelonephritis,
chronic inflammation and at last renal failure, which if left untreated, can cause death. It
also produces urease, which is responsible for the formation of bladder and kidney stones
as a result of the hydrolysis of urea to ammonia. Furthermore, the hemolysin that the
bacterium secretes is cytotoxic for urinary tract epithelial cells which are subject to invasion
by the proteus. The bacterium's swarming motility also plays an important role in renal
infections, which involve colonization of the lower urinary tract, followed by ascending
movement of the bacterium. It is believed that the swarming motility is correlated with the
effective invasion of hosts’ epithelia cells, as it provides the ability to move rapidly and
therefore to rapidly colonize and invade the cells.

The bacteria can be found throughout the kidney and bladder stones, and can reinitiate
infection after antibiotic treatment. Over time the stones may grow large enough to cause
obstruction and renal failure. Proteus can also cause wound infections, septicemia, and
pneumonias, mostly in hospitalized patients.

Introduction to Proteus mirabilis

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MIN

MAX

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

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

• P.mirabilis specific primer/probe mix (150 reactions BROWN)

FAM labelled

• P.mirabilis 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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