Executive Summary : | NTM infection is presenting a global health problem. NTM causes pulmonary and extrapulmonary disease in both immunocompetent and immunocompromised hosts. Phenotypic and molecular methods could not extensively and rapidly diagnose NTM, and drug resistance. There are many gaps in NTM knowledge, ubiquitous exposure to the microorganism, cumbersome diagnostic tests, difficulties in NTM treatment regimens, and limited data available on phenotypic and genotypic drug resistance patterns of NTM. Next-generation sequencing gives an “all-in-one” approach to drug susceptibility testing (DST) and the genotype of NTM strains. This is a prospective cross-sectional study. The study will identify the clinically relevant NTM species from clinical samples by GenoType® Mycobacterium CM/AS assay and rule out the NTM/TB mixed infection. The confirmed NTM species will be further tested to determine the phenotypic drug susceptibility of NTM isolates by micro broth dilution using Trek sensititre plates. The evaluation of the genomic pattern of drug resistance will be done by using the Whole Genome Sequencing (WGS) and exploring the correlation between clinical presentation and epidemiology of NTM in the central region of India. The clinically suspected cases of TB/NTM from whose sample will be referred to the Mycobacteriology lab for culture will be included in this study. The samples will be processed using the NALC-NaOH decontamination method, ZN staining, mycobacterial culture using the BACTEC MGIT-960 system, and Lowenstein Jensen (LJ) medium. NTM identification and differentiation will be done by GenoType Mycobacterium CM/AS assays according to the manufacturer’s instructions. The phenotypic drug susceptibility will be performed by using Trek sensititre plates for rapid and slow-growing mycobacteria according to the manufacturer’s instructions. Whole Genome Sequencing (WGS) will be performed using ion torrent technology on Ion ChefTM Instrument. Data analysis will be performed by using the Statistical Analysis System software SPSS 20.0. The statistical significance will P-values less than 0.05 will be significant. The expected outcome from this study, identify NTM to species level, phenotypic and genotypic drug-resistant patterns of NTM strains, and deep genotypic characterization of NTM by Whole Genome Sequencing. This study will help in creating awareness among the clinicians and microbiologists regarding risk factors of NTM disease and infection control practices needed in the hospital. As per best knowledge, there is no clear-cut data/study on Whole Genome Sequencing analysis of NTM strains in the central region of India. |
Co-PI: | Dr. Shashank Purwar, All India Institute Of Medical Sciences (AIIMS) Bhopal, Madhya Pradesh-462020, Dr. Alkesh Kumar Khurana, All India Institute Of Medical Sciences (AIIMS) Bhopal, Madhya Pradesh-462020, Dr. Sagar Khadanga, All India Institute Of Medical Sciences (AIIMS) Bhopal, Madhya Pradesh-462020, Dr. Jitendra Singh, All India Institute Of Medical Sciences (AIIMS) Bhopal, Madhya Pradesh-462020 |