Bead-captured Mycobacterium tuberculosis for next-generation sequencing diagnosis of uncultured tuberculosis

Excerpt

The diagnosis of pulmonary tuberculosis relying on the culture of Mycobacterium tuberculosis from respiratory tract samples and stools may be hampered by prior antibiotic treatment [1]. Direct whole-genome sequencing (WGS) of M. tuberculosis from clinical specimens may offer a suitable alternative [2‐4]. However, current protocols are not optimized for the recovery of M. tuberculosis DNA and an estimated 75% proportion of human and microbial DNA is challenging the capacity to retrieve M. tuberculosis-specific sequences [3]. Magnetic bead protocols were shown to efficiently extract M. tuberculosis mycobacteria from human sputum among all bacteria and human cells [5]. Combining magnetic bead protocol and WGS, we made real-time diagnosis of pulmonary tuberculosis in one patient receiving antibiotics which prevented culture of the causative M. tuberculosis strain. …