Difficult to Culture Bacterial Pathogens Rapidly Detected by NGS

The suspected causative pathogens could not be confirmed by routine culture in two cases, one of unconfirmed tuberculosis and one of suspected brucellosis. Gündoğdu et al. [1] successfully were able to detect the causative agents using nanopore-based metagenomic sequencing.

Peritoneal fluid and blood were rapidly processed by human DNA depletion and isolation of microbial DNA with MolYsis™ Complete5 kit (Molzym, Germany) followed by sequencing of the DNA on a MinION (Oxford Nanopore, UK) for 72 hours. The earliest runtime meeting the confident detection threshold was only after 20 minutes for Mycobacterium tuberculosis and after 30 minutes for Brucella melitensis. A high confidence level of detection at the species level was achieved within six hours

In conclusion, the molecular approach confirmed M. tuberculosis within a few hours where culture failed or shortened the turnaround time significantly where culture was positive for B. melitensis only after seven days.