PCRmax Mycobacterium tuberculosis complex Instruction Manual

IS6110 repeat region

Mycobacterium
tuberculosis complex

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Mycobacterium tuberculosis complex is a group of bacterial pathogens responsible for
causing tuberculosis (TB) in humans. They are Gram-positive, non-motile, obligate
aerobes that are related to Actinomycetes. The Mycobacteria species within this group are
characterised by 99.9% similarity at the nucleotide level and identical 16S rRNA
sequences but they may differ widely in terms of their host tropisms, phenotypes, and
pathogenicity. Some of the members of the complex infect only one host species while
others such as Mycobacterium bovis have a wide host spectrum.

Transmission generally occurs via inhalation of infected aerosols with the bacteria able to
remain in the respiratory tract for long periods of time. In the alveoli, macrophages target
the infection but some bacteria manage to grow within the macrophage, avoiding the
immune response, and subsequently causing macrophage cell death allowing bacterial
spread via the blood to the kidneys, brain, bones and lymph nodes. Bacteria within this
group divide every 15 to 20 hours which is extremely slow compared to other bacteria.
They can survive in a dry state for weeks and withstand weak disinfectants but only grow
within a host organism.

Tuberculosis is most commonly seen as an infection of the lungs resulting in pulmonary TB
which has symptoms including weight loss, fever, and loss of appetite. Extrapulmonary TB
results from the bacterial spread from the lungs and causes variable symptoms depending
on the site of infection.

Species within this group include: Mycobacterium tuberculosis, the most common cause of
tuberculosis (TB); M. africanu, a species commonly found in West Africa which causes TB
in humans; M. Canettii which also causes TB and has been found in the Horn of Africa; M.
bovis which causes tuberculosis in bovine and humans and M. microti which causes
tuberculosis in voles but has also recently been reported to infect humans.

Introduction to Mycobacterium tuberculosis complex

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MIN

MAX

The PCR Max qPCR Kit for Mycobacterium tuberculosis complex (M.
tuberculosis_complex) genomes is designed for the in vitro quantification of M.
tuberculosis_complex genomes. The kit is designed to have the broadest detection profile
possible whilst remaining specific to the M.tuberculosis_complex genome.

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

• M.tuberculosis_complex specific primer/probe mix (150 reactions BROWN)

FAM labelled

• M.tuberculosis_complex 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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