Tuberculosis (TB) is a serious, potentially fatal disease that has been a global public health issue for centuries. The growth of the causal bacterium Mycobacterium tuberculosis (M. tuberculosis) is slow (at least 6–8 weeks) when compared to other infectious bacteria, thus posing a challenge for early diagnostics. Furthermore, TB therapy has been reported to be effective when antibiotics are administered during the early stage. Therefore, accurate and rapid diagnosis of M. tuberculosis plays an important role in TB quarantine and treatment. We therefore devised an integrated microfluidic system to automatically detect live M. tuberculosis and distinguish dead bacteria. A heparin-binding hemagglutinin antibody was used to capture bacteria within 10 min. The photo-reactive dye propidium monoazide was then used to bind double-stranded DNA from dead bacteria with high affinity (20 min) such that genes of dead bacteria were not amplified by the subsequent PCR. Additionally, after 10-min of genomic DNA isolation, the final PCR step (50 min) was carried out to detect live M. tuberculosis. It is the first time that the entire bacterial detection process (including bacterial capture, propidium monoazide treatment, lysis and PCR quantification) for live M. tuberculosis could be automated within 90 min on a single chip with a limit of detection as low as 100 colony-forming units. This integrated microfluidic system is promising for point-of-care diagnostics given its automation, high speed, low cost, low sample/reagent consumption, and reduced risk of infection for front-line medical staff.
Date:
2022-08-15
Relation:
Sensors and Actuators B: Chemical. 2022 Aug 15;365:Article number 131968.
