PCRmax Streptococcus sanguinis Instruction Manual

glucosyltransferase (gtfP) gene

Streptococcus
sanguinis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Streptococcus sanguinis is a Gram-positive, non-motile cocci belonging to the viridans
group of the Streptococcus genus and posses alpha-haemolytic capabilites. The circular
genome of this bacterium is DNA based and approximately 2.4Mbp in length encoding
2274 proteins. As a part of the healthy oral flora, S. sanguinis is often found in plaque and
dental cavities but is also capable of causing subacute bacterial endocarditis if given
access to the bloodstream.

Hydrophobic interactions mediate colonisation of the tooth surface providing an anchor for
attachment of other pathogens that colonize the oral cavity and putative surface proteins
including adhesins facilitate receptor-mediated binding to host epithelial cell. Access to the
bloodstream created by open wounds such as after surgery allows the bacterium to bind to
host platelets resulting in oxidation of iron in haemoglobin.

Subacute bacterial endocarditis (SBE) has a relatively slow process of infection and, if left
untreated, can prove fatal. Most patients present with a fever that lasts several days to 2
weeks. Common symptoms include cough, shortness of breath, joint pain, diarrhea, and
abdominal pain. In around 90% of cases, heart murmurs have also been noted. High-dose
intravenous penicillin with an aminoglycoside such as gentamicin is generally given for a
minimum of four weeks to treat SBE. Patients at risk of SBE should be given antibiotics
before most medical or dental surgeries and whenever any significant skin infection occurs
to prevent the infection.

Introduction to Streptococcus sanguinis

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MIN

MAX

Our kit for S. sanguinis has been designed for the specific and exclusive in vitro
quantification of all S.sanguinis isolates and no other streptococcus species. A highly
conserved sequence within the gtfP gene has previously been shown to be a good target
sequence in other clinical real time PCR based studies (W. kim Seow et.al., 2009). 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 Streptococcus sanguinis (S.sanguinis ) genomes is designed
for the in vitro quantification of S.sanguinis genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the S.sanguinis genome.

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

• S.sanguinis specific primer/probe mix (150 reactions BROWN)

FAM labelled

• S.sanguinis 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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