Search

The major outbreak of Corona virus disease COVID-19 caused by SARS-CoV-2 had brought about 4.55 million deaths and had shaken the health care system all over the world. From the year 2020 the recovered COVID-19 patients had started to develop microbial infection, most predominantly fungal infection in which Mucormycosis gained immediate attention as it has worsen the mortality rate in humans. In the present study of 53 COVID-19 recovered patients presented with microbial infection, the analysis of frequency distribution of fungal infection preponderantly with Rhizopus oryzae, followed by Aspergillus and Candida species.

There is an urgent need for a rapid and reliable test to detect actively multiplying Mycobacterium tuberculosisdirectly from clinical specimens for an early initiation of the appropriate antituberculous treatment. This study was aimed at the optimization and application of nested reverse transcriptase–PCR (nRT–PCR) targeting the messenger RNA of the icl2, hspx, and rRNAP1genes directly from sputum specimens, and their evaluation against the culture by the BACTEC MicroMGIT mycobacterial culture system. 203 Sputum samples from clinically suspected tuberculosis patients and 30 control specimens (clinically proven viral or bacterial infections other than tuberculosis) were included in this study. The mycobacterial culture was performed by the BACTEC MicroMGIT system following the manufacturer’s instructions. The primers for nRT–PCRs targeting icl2, hspx, and rRNAP1genes were indigenously designed using the Primer-BLAST software, and optimized for sensitivity and specificity. The icl2, hspx, and rRNAP1genes were able to pick up 63.9%, 67.2%, and 58.75%, respectively, of culture-negative sputum specimens collected from clinically suspected tuberculosis patients. However, three (1.4%) were negative for nRT–PCR, but M. tuberculosis culture positive. All the 30 controls were negative for culture by the BACTEC MicroMGIT method and all three nRT–PCR. The novel nRT–PCRs targeting icl2, hspx, and rRNAP1genes developed in this study are rapid and reliable diagnostic tools to detect viable M. tuberculosisdirectly from sputum specimens. However, further study by including a larger number of sputum specimens needs to be carried out to ascertain the diagnostic utility of the novel nRT–PCRs optimized in the study.

•This was a Pilot study conducted to find out the mutations targeting rpoB, katGand inhAgene by PCR based DNA technique in M. tuberculosisisolates.•Only 37.5% of RIF-resistant isolates showed the presence of Ser531Leu mutation.•The frequency of mutations associated with INH resistance was 31.5% and 29.6% targeting katGand I gene.