PCRmax Chlamydia trachomatis Instruction Manual

Cryptic Plasmid Helicase (DnaB)
gene

Chlamydia trachomatis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Chlamydia trachomatis is an obligate, coccoid or rod shaped bacterium that is an
intracellular pathogen of humans. It is classed as a Gram-negative bacterium although it
lacks a peptidoglycan cell wall. Its genome is circular double-stranded DNA, around
1,042K nucleotides in length with an extrachromosomal plasmid of about 7.5Kbp of which
there are 7-10 copies per bacterial particle. C. trachomatis causes Chlamydia, the most
common sexually transmitted disease in the world and Trachoma, a common cause of
preventable blindness among children in developing nations.

C. trachomatis cannot survive outside of the eukaryotic cell and is transmitted through
infected bodily secretions containing the elementary body form of the bacterium. This form
is around 0.3μm in diameter and enters the host cell of mucosal membranes by self­induced endocytosis. The elementary body survives inside the endosome by avoiding
phagolysosomal fusion and produces glycogen which triggers germination into a reticulate
body. The reticulate body undergoes binary fission within the cell, the product of which is
reticulate bodies that then transform back into elementary bodies before being released
from the host cell by exocytosis.

Chlamydia infections are often asymptomatic but in other cases after one to three weeks
incubation time symptoms in males can be seen. These include pain during urination,
discharge from the penis and pain, infection or inflammation of the testicles or testicular
ducts. Treatment with a course of antibiotics usually including doxycycline and
erythromycin can eradicate the infection but as the majority of cases are asymptomatic,
treatment is not common. Delay or lack of treatment can result in pelvic inflammatory
disease (PID) in women and infertility in both sexes.

There are over 15 serovariants of C.trachomatis, four of which are implicated in ocular
infection (serovariants A, B, Ba, and C).

Introduction to Chlamydia trachomatis

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MIN

MAX

Our kit for human Chlamydia trachomatis has been designed for the specific and
exclusive in vitro quantification of this species include all serovars A, B, Ba, C, D, E, F, G,
I, K, K1, L2, L3, H, J and does not detect other closely related Chlamydia species. 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 Chlamydia trachomatis (C.trachomatis) genomes is designed
for the in vitro quantification of C.trachomatis genomes. The kit is designed to have the
broadest detection profile possible whilst remaining specific to the C.trachomatis genome.

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

FAM labelled

• C.trachomatis 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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