PCRmax Treponema pallidum Instruction Manual

polA

Treponema pallidum

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Treponema pallidum is a gram-negative bacterium of the Spirochaetes phylum. The
bacteria are approximately 6-20μm in length with a helical shape. An endoflagella found in
the periplasmic space between the outer and inner membrane enable corkscrew motility.
The bacterial genome is one double stranded DNA circular chromosome of around 1Mbp
in length that codes for over 1000 proteins. This bacterium is the parasite responsible for
causing Syphilis and is also thought to increase the likelihood of transmission of HIV.

T. pallidum acquires all of its energy through glycolosis, but relies on a host for nutrients
and therefore cannot survive in the absence of the host cells. The initial site of infection is
the epithelial cells of the genitals, although the bacterium can spread throughout the body
to any tissue or organ. The infection is a sexually transmitted disease and is spread by
contact with skin lesions or exposed mucus membranes. However, congenital syphilis is
transmitted from mother to fetus. Previous infections with T. pallidum offer no immunity,
but the infection can be treated with antibiotics such as penicillin which it has not shown
resistance to.

T. pallidum infection progresses through 4 stages if untreated: Primary; secondary; latent
and tertiary. Primary infection presents in a chancre lesion at the site where the bacteria
penetrates the epithelial cell. These indurated ulcers are clean and painless and can last
up to 5 weeks. If left untreated the second stage of the infection follows with a rash
appearing approximately 6 weeks after the initial infection, accompanied by skin lesions.
People infected may also experience tiredness, fever, headaches, sore throat, loss of
appetite and swollen glands which can last up to 6 weeks. Several months later symptoms
disappear although the bacterium is still present, this is the latent phase where
transmission rates decrease significantly. If the infection is not treated, the tertiary phase
may begin up to several years later. At this stage, syphilis can affect the bones, the
cardiovascular system and the central nervous system causing disease.

Introduction to Treponema pallidum

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MIN

MAX

This sequence has previously been established in the scientific literature as a reliable
marker for the specific detection of T. pallidum (D.Leslie et.al. 2007). The primers and
probe have 100% homology with the three principal reference sequences for T. pallidum
polA currently in NCBI database (CP000805.1 AE000520.1 U57757.1).

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

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

• T.pallidum specific primer/probe mix (150 reactions BROWN)

FAM labelled

• T.pallidum 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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