PCRmax Streptococcus pyogenes Instruction Manual

Collagen-Like Surface Protein
(Sc1A) gene

Streptococcus
pyogenes

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Streptococcus pyogenes is a Gram-positive bacterium of the Streptococcaceae family. The
cell wall of this bacterium contains A class Streptococcus antigens and so this species is
known as a Group A Streptococcus species. The genome of this bacterium is around

1.8Mbp in length and arranged in a single, circular chromosome. S. pyogenes is known to
cause human diseases from mild skin infections to Necrotising Fasciitis and other
potentially fatal systemic diseases.

S. pyogenes adheres to host epithelial cells of the respiratory system via fibronectin
binding proteins that are expressed by the bacterium. The expression of this protein,
Protein F, which is a unique property of this bacterium, is one of its virulence factors. Also,
another protein known as M protein in conjunction with a hyaluronic acid capsule allow the
bacterium to resist host cell phagocytosis. Once attached to the host cell, the bacterium
are able to invade and multiply, progressing through deeper layers of tissue, ultimately
disrupting erythrocytes.

S. pyogenes infections usually begin in the throat causing pharyngitis or mild skin
infections such as impetigo. The spread of bacteria into the lower layers of the skin and
fascia causes cellulitis and necrotizing fasciitis respectively. Some strains of S. pyogenes
release toxins that act on many different types of host cells to cause toxic shock syndrome.
Infections with S. pyogenes can usually be treated with a course of Penicillin.

Introduction to Streptococcus pyogenes

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MIN

MAX

The target sequence (sodA) has previously been shown to be a good genetic marker for
S. pyogenes in other real time PCR based studies (Rasmussen M et.al 2000). 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 pyogenes (S.pyogenes) genomes is designed
for the in vitro quantification of S.pyogenes genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the S.pyogenes genome.

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

FAM labelled

• S.pyogenes 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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