PCRmax Prevotella intermedia Instruction Manual

RNA polymerase beta-subunit
(rpoB gene)

Prevotella intermedia

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Prevotella intermedia is a Gram-negative, rod- shaped, anaerobic bacterium of the genus
Provotella. It is associated with periodontal disease and they live in the periodontal pockets
in between the teeth where they form an oral microbiota with other microbes. P. intermedia
has also been shown to be associated with endodontic infections such as root canal
infection, apical periodontitis, and periapical lesions. The genome consists of a circular
chromosome which is approximately 2700 kb long.

Prevotella intermedia growth is stimulated by an increase in gingival crevicular fluid that
results from plaque formation, usually due to poor oral hygiene. The crevicular fluid
contains nutrients such as hemin and vitamin K, which are essential for growth of this
organism. Prevotella intermedia often invade the human buccal epithelial cells. Cellular
invasion allows access to a nutritionally rich environment as well as evasion from the host
immune defences. The type C fimbriae have been shown to play a major role in the
invasion process. Prevotella intermedia also secretes salivary IgA proteases which
degrades salivary IgA1 and IgA2, allowing survival of the first line of immune defence and
access to the oral mucosa. As the bacterial load increases the gingival tissues swell, bleed
and become edematous, releasing further nutrients to promote P. intermedia proliferation.
The host launches various inflammatory responses against the bacteria, as well as
activation of matrix metalloproteases that are responsible for collagen loss in tissues. Such
collagen loss leads to attachment loss, thereby deepening the depression where gingival
tissues contact the tooth surface. This further deepening creates the periodontal pocket.
The loss of attachment is also responsible for converting gingivitis to periodontitis

Periodontitis is the disruption of connective tissue attachment along with the adjacent
alveolar bone. Some symptoms include development of new spaces between the teeth,
change in the manner that the teeth fit together when biting and swelling of the gums.
Periodontitis is significantly correlated with an overgrowth of a large number of anaerobic
bacteria so this disease can be suppressed by good oral hygiene and by utilizing
antimicrobial agents that target the participating anaerobes.

Introduction to Prevotella intermedia

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MIN

MAX

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

The primers and probe sequences in this kit have 100% homology with a broad range of
P.intermedia 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.intermedia specific primer/probe mix (150 reactions BROWN)

FAM labelled

• P.intermedia 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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