CRISPR-powered electrochemical microfluidic multiplexed biosensor for target amplification-free miRNA diagnostics

Recently the use of microRNAs (miRNAs) as biomarkers for a multitude diseases has gained substantial significance clinical well point-of-care diagnostics. Amongst other challenges, however, it holds central requirement that concentration given miRNA must be evaluated within context factors in order to unambiguously diagnose one specific disease. In terms development diagnostic methods and devices, this implies an inevitable demand multiplexing able gauge abundance several components interest patient's sample parallel. study, we design implement different multiplexed versions our electrochemical microfluidic biosensor by dividing its channel into subsections, creating four novel chip designs amplification-free simultaneous quantification up eight miRNAs on CRISPR-Biosensor X (‘X’ highlighting aspect device). We then one-step model assay followed amperometric readout combination with 2-min-stop-flow-protocol explore fluidic mechanical characteristics limitations device. The sensor showing best performance, is subsequently used Cas13a-powered proof-of-concept measurement two (miRNA-19b miRNA-20a) from miRNA-17–92 cluster, which dysregulated blood pediatric medulloblastoma patients. Quantification latter, alongside negative control measurements are accomplished same thereby confirm applicability platform challenge amplification-free, parallel detection multiple nucleic acids.