Validation of Sputum as Direct DNA Source for Whole Genome Sequencing of Mycobacterium Tuberculosis

This study was terminated on September 27, 2021

Dr. Jeffrey Tornheim is Principal Investigator for this study. 

 There are limited studies available from India on whole genome sequencing (WGS).  This is a first-of-its-kind wherein WGS for MTB will be carried out utilizing direct sputum DNA and simultaneously look on the cultured DNA from the paired samples. Achieving this would revolutionize the management of TB with the introduction of WGS as a diagnostic test and pave the way for personalized treatment. With a minimal turnaround time, WGS is likely to be much faster than traditional phenotyping and could challenge molecular tests such as Xpert MTB/RIF which is the current rapid diagnostic test. 

Objectives:

• Validate the DNA extracted from direct sputum samples for WGS 
• WGS in the DNA samples obtained from the paired cultures of these sputum samples
• Validation of 2 extraction kits to quantify the DNA from the sputum samples
• The library preparation will also be carried out in parallel using the random universal primer and Sure select Primer designed specifically to the standard H37Rv Laboratory strain