Rapid Tuberculosis Diagnostics Including Molecular First- and Second-Line Resistance Testing Based on a Novel Microfluidic DNA Extraction Cartridge

Two-stage tuberculosis diagnostics: combining centrifugal microfluidics to detect TB infection and Inh and Rif resistance at the point of care with subsequent antibiotic resistance profiling by targeted NGS

“…For the identification of the analytical sensitivity, the pathogenic MTBC strain H37Rv (ATCC, Manassas, VA;35829) was spiked in TB negative tested human sputum with real colony count correction of the bacteria concentration similar to previous studies. 20,40…”

“…For the identification of the analytical sensitivity, the pathogenic MTBC strain H37Rv (ATCC, Manassas, VA;35829) was spiked in TB negative tested human sputum with real colony count correction of the bacteria concentration similar to previous studies. 20,40 Diagnostic sensitivity and specificity. The diagnostic sensitivity was determined with bacteriologically confirmed, randomly selected, microscopically positive sputum samples from TB patients in Nepal at the German Nepal TB Project (GENETUP) in Kalimati, Kathmandu-Valley, Nepal.…”

“…Automated centrifugal microfluidic sample preparation of DNA from clinical sputum samples was shown previously. 19,20 With this approach, detection of MTBC and resistance markers must subsequently be performed in manual steps and with an additional PCR cycler. This limits its application at the PoC, as a laboratory infrastructure is required.…”

“…This limits its application at the PoC, as a laboratory infrastructure is required. Here, the previously demonstrated centrifugal microfluidic purification of DNA from clinical sputum samples for subsequent molecular TB diagnostics 19,20 is further developed to a fully integrated sample-to-answer analysis. Automated PoC sample-to-answer analysis for pathogen detection is in principle not new to the field.…”

Microfluidic systems for infectious disease diagnostics

“…The availability of two identical DNA extracts significantly increased the versatility of the diagnostic workflow. 215 Kaur et al proposed a low-cost hybrid paper/plastic device for LAMP-based TB assays on 12 test zones. The chip features an array of sealed paper pads as amplification sites with dry-stored reagents and fluorescence smartphone read-out.…”