Mohd Nawi, Mohd Amiruddin
(2015)
Development of mycobacterium tuberculosis 16 kDa protein detection using immunochromatography test.
Masters thesis, Universiti Sains Malaysia.
Abstract
Mycobacterium tuberculosis (Mtb) is a pathogenic bacterium that can cause
tuberculosis (TB). This contagious disease remains a severe health problem in the
world. The disease is transmitted via inhalation of airborne droplets carrying Mtb
from TB patients. Early detection of infection is vital to prevent transmission of the
disease to people in close contact with the patients. To date, there is no simple, rapid,
sensitive and specific diagnostic test for TB. Previous studies showed the potential of
Mtb 16 kDa antigen in TB diagnosis. In this study, an immunochromatographic
(ICT) lateral flow dipstick assay against 16 kDa antigen was developed and assessed
as a potential rapid method to diagnose TB. A 16 kDa monoclonal antibody was
optimized as the capturing and detection antibody on the ICT platform. The
parameters of control line, blocking reagent, running buffer, conjugate and sample
pad were also optimized before a complete prototype of TB-antigen ICT dipstick was
developed. Sensitivity studies on TB-antigen ICT prototype dipstick showed that the
dipstick was capable to detect purified 16 kDa antigen as low as 125 ng. However,
the specificity of the assay was inconclusive probably due to the method used for
sample preparation, which needs further study. The analytical sensitivity suggests its
potential usefulness in different clinical applications.
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