Simple Polydisperse Droplet Emulsion Polymerase Chain Reaction with Statistical Volumetric Correction Compared with Microfluidic Droplet Digital Polymerase Chain Reaction.

摘要: Nucleic acid amplification technology, such as polymerase chain reaction (PCR), has enabled highly sensitive and specific disease detection quantification, leading to more accurate diagnosis treatment regimens. Lab-on-a-chip applications have developed methods partition single biomolecules, DNA RNA, into picoliter-sized droplets. These individual vessels lead digitization of PCR enabling improved time direct quantification nucleic acids without a standard curve, therefore simplifying assay analysis. Though impactful, these improvements generally been restricted centralized laboratories with trained personnel expensive equipment. To address limitations make this technology applicable for variety settings, we statistical framework apply droplet performed in polydisperse droplets prepared any specialized The system allows digital (ddPCR) reverse transcriptase (RT-ddPCR) that is comparable commercially available systems BioRad's ddPCR. Additionally, approach compatible range input sample volumes, extending the dynamic beyond commercial ddPCR systems. In work, show assays can reduce overall while still providing quantitative results. We also report multiplexed demonstrate proof-of-concept rapid preparation multiple samples simultaneously. Our simple be extended settings complex samples.