PCRmax Streptococcus mitis Instruction Manual

Streptococcus mitis phage SM1,
platelet binding protein (pb1A)

Streptococcus mitis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Streptococcus mitis are gram positive, non-motile, mesophilic bacteria of the viridan
streptococci (VS) group. They usually occur in small chains and are part of the normal oral
flora of mammals. The S. mitis genome consists of 2Mb of DNA in a circular conformation
consisting of over 2000 protein coding genes. This bacterium is usually non-pathogenic but
can cause dental infections, strep throat and infective endocarditis in susceptible
individuals.

The bacteria gain access to the blood stream during oral operative procedures and bind to
platelets most likely via bacterial derived lipoproteins Pb1A and Pb1B which target the host
receptor α2-8-linked sialic acid of ganglioside GD3 present in the platelet membrane. The
alpha-haemolytic properties of this bacterium result in oxidation of the iron in haemoglobin
causing the breakdown of red blood cells.

Infective endocarditis presents with non-specific symptoms including mild fever which if left
untreated is fatal. S.mitis infections are usually treated with single or combination
antibiotics, particularly penicillin. Infections progressing to congestive heart failure resulting
from recurring emboli often require surgery to replace the damaged valves.

Introduction to Streptococcus mitis

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MIN

MAX

The logenic phage SM1 encodes critical proteins which are required for binding of S.mitis
to human platelets. The SM1 phage is therefore predicted to be present in all cases of
human infection. Environmental samples of S.mitis may not include this sequence.

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

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

FAM labelled

• S.mitis 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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