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Light Cycler System

for MTB Qualitative


Detection
The mycobacteria are acid-fast, non motile, road-shaped,
aerobic bacteria that do not form spores. They contain several
species pathogenic to humans. Many mycobacterial species are
soil and water saprophytes. The primary human pathogens are
those species of the M.tuberculosis complex (M.tuberculosis,
M.bovis, M. africanum and M.microti), and M.leprae.
M.tuberculosis is the most important mycobacterial pathogen.
M.tuberculosis is only found in humans, who provide its
reservoir and and transmit the tubercle bacilli from person-to-
person by droplet infection. Approximately one-third of the
world’s population harbors M.tuberculosis and is at risk of
developing active tuberculosis infection. This bacteria can
cause chronic pulmonary disease but can also infect any organ
in the body. Due to the risk of the spread of the disease, the
potential for the emergence of drug-resistant strains, and the
severity of the disease in patients infected with HIV-1, the rapid
diagnosis of the M.tuberculosis complex is extremely important.

The isolation of an organism from M.tuberculosis complex is


required for the definitive diagnosis of tuberculosis. Routine
cultures are time-consuming and can take up to eight weeks.
Microscopic examination of acid-fast smears is the most rapid
method for the detection of mycobacteria, but it is insensitive
and non-specific. Immunological ans serological techniques are
limited, in general due to poor sensitivity and/or specifity.
Species-specific nucleic acid probes have significantly improved
the opportunity for rapid confirmation of culture results for
several mycobacterial species. The development of PCR-based
tests specific for mycobacteria has been shown to further
improve the rapid diagnosis of tuberculosis by allowing the

Reagents
Information

Biological Sample: Sputum, Brochial Alveolar Lavages (BAL)


Biological sample extraction kit
High Pure PCR Template Preparation Kit (Cat. No. 11 796828001)

Thermal Cycler: Light Cycler type 2.0

Amplification Specific Reagents and PCR protocols


LightCycler FastStart DNA Master Plus SYBR Green I (Cat. No. 03 515
869 001)

Primer: IS 6110 repetitive sequence of MTB


Mtb Qualitative Detection
Workflow
Manual Real Time PCR
Sputum or Decontaminati sample for Mtb DNA
BAL Swab on preparation
HighPure
PCR
Template
Preparation

PCR
Workflow
Add DNA
Template

- Reaction
Buffer
- dNTP’s
- Taq DNA Reaction Capillary
Polymerase on LightCycler 2.0
- Water PCR and analysis
Grade

Result
s

PT Roche Indonesia Tel. 21 30413000


Diagnostics Division Fax. 21 51400090
Artha Graha Lt.21-SCBD Lot 25 Email: lianita.salim@roche.com
Jl. Jend.Sudirman Kav.52-53
Jakarta 12190 ratna_agung_aman.taufani@roche.com

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