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- International journal of dermatology 01/2015; 54(1). DOI:10.1111/ijd.12636
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ABSTRACT: : Keto-iminol transformation of formohydroxamic acid in the presence of water molecules is investigated using ab initio quantum mechanics calculations. Calculations revealed that the proton transfer energy barriers are significantly lowered in the presence of water molecules. The proton transfer in Z-keto form occurs via a pathway involving two transition states and a charge separated intermediate. The proton transfer in E-keto tautomer occurs via a single transition state. A third pathway, which is not feasible in the absence of water, becomes feasible when there are hydrogen bonded water molecules. The stability order of formohydroxamic acid tautomers was found to be E-keto > Z-keto > Z-iminol > E-iminol and remains the same even after hydrogen bonded to water.Computational and Theoretical Chemistry 03/2014; 1032. DOI:10.1016/j.comptc.2014.01.027 - [Show abstract] [Hide abstract]
ABSTRACT: Rifampicin resistance of Mycobacterium tuberculosis is due to the occurrence of point mutations of the rpoB gene and the site of mutations vary geographically. Commercialized molecular based methods are not able to comprehensively detect rifampicin resistance as they target a limited number of gene mutations which are thought to be common. The aim of the study was to establish a low cost DNA probe based colorimetric method that can be customized for detection of rifampicin resistance of M. tuberculosis. Thus, Enzyme-linked oligosorbent assay (ELOSA) was developed for the detection of polymerase chain reaction (PCR) amplified fragments of rpoB gene of M. tuberculosis DNA on microtiter plates. Forty two M. tuberculosis isolates (rifampicin resistant and susceptible isolates identified by agar proportion method) were used for developing and validating the assay. The point mutations of resistant isolates had been previously determined by DNA sequencing. Two fragments of rpoB gene were labeled with digoxigenin by PCR. The amplified products were hybridized with selected allele specific probes for three mutations and its wild types (six probes) which were captured onto streptavidin coated microtiter plates and was detected by colour development. Both sensitivity and specificity of all probes were≥96% and there was excellent discrimination (area under the curve (AUC)>0.9) between rifampicin susceptible cases and resistant cases. The probe-based colourimetric assay (PCR-ELOSA) developed in this study showed good agreement with reference mutation that were confirmed by DNA sequencing. In conclusion, PCR-ELOSA is a reliable and economical assay that can be customized for detection of rifampicin resistance.Journal of microbiological methods 11/2013; 96(1). DOI:10.1016/j.mimet.2013.11.006
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