PCRmax Treponema denticola Instruction Manual

Cytoplasmic filament protein A
(CfpA) gene

Treponema denticola

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Within the oral cavity, treponemes are most often associated with diseases of the
periodontium such as early-onset periodontitis, necrotizing ulcerative gingivitis, and acute
pericoronitis. T.denticola has a 2.84 Mbp single circular chromosome containing approx
~2,770 predicted protein-encoding genes which is considerably larger in size than the
related syphilis causing Treponema pallidum.

It primarily utilizes serine, alanine, cysteine, and glycine when grown in vitro and generates
fermentation products. These metabolites can accumulate in high concentrations and
could influence the composition of bacterial species within accumulated plaques or biofilms
as well as affecting host tissues in the surrounding areas.

Part of the natural oral flora T.denticola can inflame the gingeva leading to gingivitis and
then ulcerative gingivitis requiring medical-dental intervention. Transmission is usually a
non-issue due to natural oral flora makeup. Main risk factors are weakened immune
system and poor oral health.

Some of the characteristics of T. denticola that represent its major virulence factors in
chronic periodontitis are its motility and chemotaxis, which enable the bacterium to rapidly
colonize new sites, penetrate deep periodontal pockets, and penetrate epithelial layers as
well as its ability to form biofilms and a range of cell-surface proteins to overcome the host
defence in order to protect the subgingival biofilm and cause tissue destruction.

Introduction to Treponema denticola

2

MIN

MAX

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

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

3

Kit Contents

• T.denticola specific primer/probe mix (150 reactions BROWN)

FAM labelled

• T.denticola 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

4