Targeted sputum sequencing for rapid and broad drug resistance of Mycobacterium tuberculosis.

ABSTRACT:

Purpose

Rapid detection of drug resistance in Mycobacterium tuberculosis (Mtb) from clinical samples facilitates the timely provision of optimal treatment regimens for tuberculosis (TB) patients.

Methods

In November, 2023, the WHO released its second catalogue of resistance-conferring mutations in Mtb. Utilizing this information, we developed a single 17-plex PCR assay covering 16 key resistance genes and modified thermo-protection buffer to amplify 30 kbp DNA directly from sputum samples for nanopore sequencing. We implemented our protocol using rapid barcoding for sequencing with both a Flongle and a MinION flow cell.

Results

The single multiplex PCR assay was successfully validated on clinical sputum samples using the thermo-protection buffer. The protocol was appl

SUBMITTER: Dou HY 

PROVIDER: S-EPMC12316706 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature