Sample Preparation and qPCR Detection of Tuberculosis on a Centrifugal Microfluidic Cartridge Enabling Molecular Downstream Resistance Profiling

Tuberculosis (TB) is still one of the world’s deadliest infections. Fast detection of the pathogen M. tuberculosis (MTB) and its genetic resistance markers substantially improves treatment success and outcome. A key element for rapid genetic diagnostics is the efficient extraction of DNA from sputum for qPCR detection at the point of care. We present the fully automated sample preparation of MTB DNA from 3 mL of liquefied sputum and qPCR detection of MTB on a centrifugal microfluidic cartridge. Our method achieves a limit of detection (LoD) between 17 and 57 CFU/mL and provides a purified DNA solution for molecular downstream testing, such as targeted NGS.