Rapid detection of Mycobacterium tuberculosis including drug resistant strains and Melioidosis

Abstract

Tuberculosis remains a leading health concern worldwide. Melioidosis mimicker of Tuberculosis, may be misdiagnosed as Tuberculosis. We aimed to detect Mycobacterium tuberculosis using Polymerase chain reaction drug resistant mutations using sequencing and Burkholderia pseudomallei using nested PCR. Ethics committee approval was obtained. Microscopy, culture and uniplex, duplex, triplex PCR targeting IS6110 TRC4 and/or MPB64 were performed for MTB detection. rpoB katG genes by Sangers and whole genome by NGS were sequenced. B.pseudomallei specific 16S 23S spacer region was targeted by nested PCR newlineTotally 217 patients were included Group 1 106 pulmonary and Group 2 68 pulmonary 43 extra pulmonary. The sensitivity of PCR increased by using TRC4 100 per cent The specificity remained 100 per cent for IS6110 duplex/triplex Mutations conferring MDR were found in 6 50 per cent strains. The WGS/NGS analysis revealed one MDR to be Pre XDR TB Among 125 samples tested, 8 per cent were found positive by nested PCR for B pseudomallei Patients presented clinical manifestations similar to TB and co-morbidities CKD malignancy diabetes mellitus, HIV and alcoholism Additional targets in PCR help in rapid detection of MTB NGS rapidly identified a Pre XDR and confirmed other MDR strains It is vital to screen for B pseudomallei in patients suspected for TB newline newline newline