PCRmax Aggregatibacter actinomycetemcomitans Instruction Manual
hemoglobin-bindingprotein
(hbpA) gene
Aggregatibacter
actinomycetemcomitan
s
PCRmax Ltd qPCR test
TM
150 tests
For general laboratory and research use only
1
Aggregatibacter actinomycetemcomitans is a non-motile, rod-shaped, Gram-negative
bacterium. This bacterium is also a facultative anaerobe which can grow under aerobic or
anaerobic conditions. The 43Kbp genome is arranged in a circular configuration and
contains up to 60 open reading frames. The outer surface of the cell includes appendages
known as fimbriae which are used for host cell adhesion.
A. actinomycetemcomitans inhabits the human oral cavity where it adheres to the mucosal
tissue of the gum using fimbriae. Here, they are able to create a biofilm with other resident
bacterial species that if not treated can cause periodontitis or gingivitis.
The bacterium produces leukotoxin which can be found on the outer bacterial membrane
and is used to avoid the host cell immune response. This evasion allows the bacteria to
stimulate a chronic inflammatory response that leads to damage of the periodontium which
includes connective tissue, ligaments and bone.
The damage to the periodontium that is caused by A. actinomycetemcomitans results in
redness, bleeding or swelling of the gums which progresses to recession of the gums that
can lead to deep pockets forming between the teeth when the attached gingiva is
destroyed. Eventually the teeth will become lose and fall out.
Introduction to Aggregatibacter
actinomycetemcomitans
2
MIN
MAX
The haemoglobin binding protein gene has previously been identified as a highly specific
market for A. actinomycetemcomitans (J. Periodont Res 2008; 43: 352-359). The primers
and probe have 100% homology with all reference sequences for A.
actinomycetemcomitans in the NCBI database.
EF142372.1, EF142371.1, EF142368.1, EF142367.1, EF142366.1, EF142364.1,
EF142363.1, AF359451.1, AF359444.1, AF359442.1, EF142443.1, EF142442.1,
EF142441.1, EF142440.1, EF142439.1, EF142438.1, EF142437.1, EF142436.1,
EF142435.1, EF142434.1, EF142433.1, EF142432.1, EF142431.1, EF142430.1,
EF142429.1, EF142428.1, EF142427.1, EF142426.1, EF142425.1, EF142424.1,
EF142423.1, EF142422.1, EF142421.1, EF142420.1, EF142419.1, EF142418.1,
EF142417.1, EF142416.1, EF142415.1, EF142414.1, EF142413.1, EF142412.1,
EF142411.1, EF142410.1, EF142409.1, EF142408.1, EF142407.1, EF142406.1,
EF142405.1, EF142404.1, EF142403.1, EF142402.1, EF142401.1, EF142400.1,
EF142399.1, EF142398.1, EF142397.1, EF142396.1, EF142395.1, EF142394.1,
EF142393.1, EF142392.1, EF142391.1, EF142390.1, EF142389.1, EF142388.1,
EF142387.1, EF142386.1, EF142385.1, EF142384.1, EF142383.1, EF142382.1,
EF142381.1, EF142380.1, EF142379.1, EF142378.1, EF142377.1, EF142376.1,
The PCR Max qPCR Kit for Aggregatibacter actinomycetemcomitans (A.actino) genomes
is designed for the in vitro quantification of A.actino genomes. The kit is designed to have
the broadest detection profile possible whilst remaining specific to the A.actino genome.
The primers and probe sequences in this kit have 100% homology with a broad range of
A.actino 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
3
Kit Contents
• A.actino specific primer/probe mix (150 reactions BROWN)
FAM labelled
• A.actino 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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