PCRmax Filifactor alocis Instruction Manual

DNA gyrase subunit B (gyrB)
gene

Filifactor alocis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Filifactor Alocis is a bacteria which can develop complex communities that attach to the
tooth surface. These highly dynamic microfloral environments are challenging both
clinicians due to F. alocis being a fastidious, Gram-positive, obligate anaerobic rod that
possesses trypsin-like enzymatic activity. In this way it is similar to other detrimental oral
bacteria such as P. gingivalis and T. denticola which can also cause periodontal. Because
of this, F. Alocis presence may be utilised as a marker for periodontal deterioration.

F Alocis contains approx. 1.93mbp which can code for ~1650 genes. It is a source of
periodontitis in humans.

Periodontitis is a chronic inflammatory bacterial infection leading to destruction of
periodontal ligaments (i.e. the tissues that surround and support the tooth) and the
supporting bone of the tooth. Periodontal pockets accommodate a multitude of bacterial
types. The organisms do not live in a planktonic form, but rather as a sessile community
attached to the tooth surface in a matrix of extracellular polymers.

Periodontitis involves progressive loss of the alveolar bone around the teeth, and if left
untreated, can lead to the loosening and subsequent loss of teeth. Periodontitis is caused
by microorganisms that adhere to and grow on the tooth's surfaces and the subsequent
immune response against these microorganisms. A diagnosis of periodontitis is
established by inspecting the soft gum tissues around the teeth in a clinical examination
and by evaluating the amount of bone loss around the teeth.

Periodontitis beyond the gingivitis stage is destructive (i.e. irreversible) and may affect
different sites around the tooth area. Severity of periodontitis is determined by the amount
of ligament lost to the disease resulting in loosening/detachment

Symptons include: Redness or bleeding of gums while brushing teeth, using dental floss or
biting into hard food (e.g. apples), reoccurring gum swelling, halitosis, metallic taste in
mouth, recession of gums and loose teeth. Gingival inflammation and bone destruction are
largely painless and so may go unnoticed for a lengthy time.

Periodontitis has been linked to increased inflammation in the body, such as indicated by
raised levels of C-reactive protein and interleukin-6 as well as being linked through this to
increased risk of stroke and myocardial infarction.

Treatment involves extensive ongoing oral hygiene as well as ongoing care by a dentist.

Introduction to Filifactor alocis

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MIN

MAX

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

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

• F.alocis specific primer/probe mix (150 reactions BROWN)

FAM labelled

• F.alocis 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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