PCRmax Chlamydiaceae (all species) Instruction Manual

16s ribosomal RNA sequence

Chlamydiaceae (all
species)

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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The bacterial family, Chlamydiaceae, contains two genera of closely related, Gram­negative pathogens. They are all aerobic, non-motile bacterium with a coccoid or rod­shaped cell body. The genus Chlamydia contains three species: Chlamydia muridarum, C.
suis and C. trachomatis. The latter of which infects humans while the other species infect
mice and swine respectively. The second genus, Chlamydophila represents the species
Chlamydophila pneumoniae, C. pecorum, C. psittaci, C. abortus, C. felis and C. caviae. All
species of the Chlamydiaceae family are around 0.1µm-0.8µm in length with genomes of
approximately 1Mbp length in a circular arrangement usually with one plasmid.

All species within this family are obligate pathogens, and are metabolically inert outside of
a host. Upon infection of a host, the dispersal form of the bacterium, known as elementary
bodies, attach to the host cells and are subsequently engulfed by phagocytosis. Once
within the cell, the elementary bodies transform into reticulate bodies which undergo binary
fission in the host cells’ inclusion bodies to gives rise to new elementary bodies which are
then released by cell lysis allowing for spread and infection of new host cells.

Each species target specific hosts and can cause a range of illnesses that vary between
host species.
The most commonly known species is C. trachomatis, the causative agent of Chlamydia,
the sexually transmitted disease. This bacterium can also be transmitted from mother to
child during pregnancy and infect the eyes causing conjunctivitis. The genital infection
causes urethritis, epididymitis and prostatitis in males and Pelvic Inflammatory Disease
(PID) in females with an increased risk of contracting HIV. Transmission of the bacteria
occurs via contact with infected bodily fluids which then infect mucosal membranes. The
infection can be treated with a course of antibiotics.
Sexually transmitted infections in females are most often asymptomatic, but can manifest
in chronic pain of the pelvic region, vaginal bleeding and painful urination. Infection of the
ovaries, fallopian tubes or uterus causes Pelvic Inflammatory Disease (PID) which can
lead to difficulties in conceiving, increased risk of ectopic pregnancy or infertility. Infections
in males are more likely to be symptomatic, causing painful urination, discharge from the
penis and swollen testicles and may eventually causes infertility if left untreated.
Eye infections that cause conjunctivitis or trachomas are thought to cause up to 15% of
cases of blindness and C. trachomatis is therefore an important pathogen in this area.

Chlamydia muridarum causes pharyngitis, bronchitis, and pneumonitis in infected mice and
hamsters and is most likely to be transmitted via contact with infected bodily fluids. While
Chlamydia suis causes enteritis, pneumonia and conjunctivitis in infected swine and is
likely to be transmitted in the same way.

Introduction to Chlamydiaceae (all species)

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MIN

MAX

The primer and probe sequences have 100% homology to C. muridarum, C.suis, C.
trachomatis, C.pneumonia, C.pecorum, C.abortus, C.psittaci, and C.caviae 16S
ribosomal RNA sequences.

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

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

• Chlamydia specific primer/probe mix (150 reactions BROWN)

FAM labelled

• Chlamydia 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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