PCRmax Clostridium tetani Instruction Manual

tetanus neurotoxin heavy chain 2

Clostridium tetani

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Clostridium tetani is a rod-shaped, Gram-positive, anaerobic bacterium which occurs
naturally in soil or as a parasite in the gastrointestinal tract of animals. The genome is
2,799,250 bp in length arranged as a double-stranded circular DNA molecule with a
plasmid of over 74K bp encoding an exotoxin. C.tetani is responsible for causing tetanus in
humans and horses; a disease characterised by painful muscular spasms that can lead to
respiratory failure.

C. tetani enters the host through open wounds and once inside the body begins to
replicate. It produces an exotoxin called the tetanospasmin which is released during spore
germination. This toxin blocks neurotransmitter release from the presynaptic membrane of
inhibitory interneurons of the spinal cord and brainstem that regulate muscle contraction.
This interference in the normal nerve functioning causes continuous muscle contraction.
The tetanus infection can be either localised or general with the former being confined to
the peripheral nerves and the latter passing through the blood and lymph to the motor
nerves.

Generalised infections most commonly occur in the face and neck, causing trismus or
lockjaw. Continuous activation of the motor nerves causes extensive muscle cramping and
twitching leading to spasms moving from the neck to the trunk and limbs which, if left
untreated, may result in respiratory failure following obstruction of the upper airway. Since
the introduction of a vaccine in the 1920s, C.tetani infections in industrialised countries
have been dramatically reduced with cases most commonly caused by insufficient
immunisation.

Introduction to Clostridium tetani

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MIN

MAX

Our kit for C. tetani has been designed for the specific and exclusive in vitro quantification
of this species. The Clostridium tetani toxin gene is unique to this species and other
clostridium within this genus are not detected. 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 Clostridium tetani (C.tetani) genomes is designed for the in
vitro quantification of C.tetani genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the C.tetani genome.

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

• C.tetani specific primer/probe mix (150 reactions BROWN)

FAM labelled

• C.tetani 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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