ABSTRACT
A 3D printed (3DP) microfluidic polymerase chain reaction (PCR) device was demonstrated by detecting synthetic SARSCoV-2 at 106 copies/μL. The microfluidic device was fabricated using stereolithography 3DP and had a reaction volume of ~22 nL. The microdevice showed PCR amplification with 85 base synthetic ssDNA targets and primers designed for a SARS-CoV-2-specific region. The device was 2.5 times faster compared to a qPCR instrument with >60,000 times smaller reagent volume. The 3DP microdevice is a promising technology to significantly reduce the manufacturing costs of microfluidic devices that could be used towards point-of-care applications. © 2023 SPIE.
ABSTRACT
We will present a microfluidic assay to detect SARS-CoV-2 RNA from nasopharyngeal swab samples. Our method leverages isotachophoresis (ITP) to integrate sample preparation, RT-LAMP, and CRISPR-based nucleic acid detection in an automatable chip. For the first time, we use ITP to purify, pre-concentrate and isothermally amplify target nucleic acids into a ~1 µL reaction volume on-chip. The device then transitions LAMP amplicons into an on-chip zone containing Cas12-gRNA complexes and reporter molecules to measure target-activated CRISPR activity. We will use our method to automatically detect COVID-19 from nasopharyngeal swab samples. © 2021 MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. All rights reserved.