Development Of A Nucleic Acid Amplification Test For The Detection Of Mycobacterium Tuberculosis
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Date
2015-10
Authors
Kaniappan, Priyatharisni
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Tuberculosis is one of the common infectious diseases in Malaysia, caused by
the airborne pathogen, Mycobacterium tuberculosis. Due to the infectious nature of the
disease, diagnostic-based detection of Mycobacterium tuberculosis is vital for effective
disease management, surveillance, epidemiological purpose and for the administration of
a prompt treatment. Current diagnostic methods rely on smear microscopy and culture
method; however these strategies suffer from several pitfalls. This shifts the attention
towards the nucleic-acid based detection of M. tuberculosis, which requires the
identification of suitable nucleic acid target as the diagnostic marker. One novel class of
nucleic acid, which is the non-protein-coding RNAs, is a class of RNA that is not
translated into protein but perform certain regulatory functions, suggesting the
potentiality to be harnessed as a diagnostic biomarker. Bioinformatic-based analysis
revealed the presence of 140 npcRNA genes, one of it, npcTB 6715, which is specific
for M. tuberculosis is chosen as the potential diagnostic marker. As the most established
nucleic acid-based detection platform, PCR assay is chosen as the platform of choice to
test the potentiality of this npcRNA for the diagnostic-based detection of
M. tuberculosis. The PCR assay developed also incorporates IS6110 as the counterpart
of npcTB 6715, whereby the usage of the npcRNA is able to detect the IS6110-deficient
strains of M. tuberculosis, averting false-negative results. The multiplex PCR assay
developed showed a detection limit of 10 pg. Evaluation with 500 Mycobacterium
cultures demonstrated values of 98.4%, 96.1%, 98.6% and 95.4%, respectively for the
sensitivity, specificity, Positive Predictive Value and the Negative Predictive Value. The
values obtained suggested the diagnostic potentiality of the mPCR, as a promising
ASSURED’ diagnostic test for the specific detection of M. tuberculosis and
corroborated the diagnostic potentiality of the npcTB 6715.
Description
Keywords
Development of a nucleic acid amplification , detection of mycobacterium tuberculosis