A sample-in-digital-answer-out system for rapid detection and quantitation of infectious pathogens in bodily fluids

摘要: A variety of automated sample-in-answer-out systems for in vitro molecular diagnostics have been presented and even commercialized. Although efficient operation, they are incapable quantifying targets, since quantitation based on analog analytical methods (via standard curve analysis) is complex, expensive, challenging. To address this issue, herein, we describe an integrated sample-in-digital-answer-out (SIDAO) diagnostic system incorporating DNA extraction digital recombinase polymerase amplification, which enables rapid quantitative nucleic acid analysis from bodily fluids within a disposable cartridge. Inside the cartridge, reagents pre-stored sterilized tubes, with pipetting module allowing facile liquid transfer. For analysis, fabricate simple, single-layer polydimethylsiloxane microfluidic device develop novel simple sample compartmentalization strategy. Sample solution partitioned into array 40,044 fL-volume microwells by sealing through application mechanical pressure. The entire performed portable, fully instrument. We evaluate capabilities analyzing Mycobacterium tuberculosis genomic both spiked saliva serum samples, demonstrate excellent accuracy specificity. This SIDAO provides promising platform testing at point-of-care.