PCRmax Pseudomonas aeruginosa Instruction Manual
RegA (toxin A synthesis
regulating gene)
Pseudomonas
aeruginosa
PCRmax Ltd qPCR test
TM
150 tests
For general laboratory and research use only
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Pseudomonas aeruginosa is a Gram-negative, aerobic, rod-shaped bacterium with
unipolar motility. An opportunistic human pathogen, P. aeruginosa is also an opportunistic
pathogen of plants. P. aeruginosa is the type species of the genus Pseudomonas (Migula
1894).
P. aeruginosa secretes a variety of pigments, including pyocyanin (blue-green), fluorescein
(yellow-green and fluorescent, now also known as pyoverdin), and pyorubin (red-brown).
King, Ward, and Raney developed Pseudomonas Agar P (aka King A media) for
enhancing pyocyanin and pyorubin production and Pseudomonas Agar F (aka King B
media) for enhancing fluorescein production.
P. aeruginosa is often preliminarily identified by its pearlescent appearance and grape-like
odor in vitro. Definitive clinical identification of P. aeruginosa often includes identifying the
production of both pyocyanin and fluorescein as well as its ability to grow at 42°C. P.
aeruginosa is capable of growth in diesel and jet fuel, where it is known as a hydrocarbon
utilizing microorganism, causing microbial corrosion. It creates dark gellish mats
sometimes improperly called "algae" because of their appearance.
Although classified as an aerobic organism, P. aeruginosa is considered by many as a
facultative anaerobe as it is well adapted to proliferate in conditions of partial or total
oxygen depletion. This organism can achieve anaerobic growth with nitrate as a terminal
electron acceptor, and in its absence it is also able to ferment arginine by substrate-level
phosphorylation. Adaptation to microaerobic or anaerobic environments is essential for
certain lifestyles of P. aeruginosa, like during lung infection in cystic fibrosis patients where
thick layers of alginate surrounding bacterial mucoid cells can limit the diffusion of oxygen.
Introduction to Pseudomonas aeruginosa
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MIN
MAX
The target sequence (regA) has previously been shown to be a good genetic marker for
H. influenza in other real time PCR based studies (K.E. Shannon et.al. 2008). The
primers and probe sequences in this kit have 100% homology with a broad range of
clinically relevant reference sequences based on a comprehensive bioinformatics
analysis.
The PCR Max qPCR Kit for Pseudomonas aeruginosa (P.aeruginosa) genomes is
designed for the in vitro quantification of P.aeruginosa genomes. The kit is designed to
have the broadest detection profile possible whilst remaining specific to the P.aeruginosa
genome.
The primers and probe sequences in this kit have 100% homology with a broad range of
P.aeruginosa 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 help@pcrmax.com and our bioinformatics team will
answer your question.
Specificity
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Kit Contents
• P.aeruginosa specific primer/probe mix (150 reactions BROWN)
FAM labelled
• P.aeruginosa 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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