AN INTEGRATED RT-LAMP AND CRISPR ASSAY FOR NUCLEIC ACID DETECTION IN A SINGLE MICROFLUIDIC CHIP

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.

A microfluidic approach to rapid crispr-based detection of sars-cov-2 RNA

We will present a microfluidic assay to detect SARS-CoV-2 RNA from raw nasopharyngeal swab samples in <30 min. Our method combines isotachophoresis (ITP) and state-of-the-art CRISPR Cas12-based nucleic acid detection. We show, for the first time, that electric field gradients in ITP can be used to pre-concentrate Cas12-guide RNA complex, target nucleic acids, and reporter molecules by ~1,000-fold into a ~100 pL volume on-chip. This pre-concentration accelerates target-activated CRISPR enzymatic activity on reporter molecules. We combine our ITP-CRISPR method with ITP-based nucleic acid extraction and isothermal amplification to detect Covid-19 from both contrived and clinical patient samples. © 2020 CBMS-0001