1. Point-of-care CRISPR-Cas-assisted SARS-CoV-2 detection in an automated and portable droplet magnetofluidic device.
- Author
-
Chen, Fan-En, Lee, Pei-Wei, Trick, Alexander Y., Park, Joon Soo, Chen, Liben, Shah, Kushagra, Mostafa, Heba, Carroll, Karen C., Hsieh, Kuangwen, and Wang, Tza-Huei
- Subjects
- *
SARS-CoV-2 , *MICROFLUIDIC devices , *VIRUS diseases , *COVID-19 , *INFECTIOUS disease transmission , *NUCLEIC acids - Abstract
In the fight against COVID-19, there remains an unmet need for point-of-care (POC) diagnostic testing tools that can rapidly and sensitively detect the causative SARS-CoV-2 virus to control disease transmission and improve patient management. Emerging CRISPR-Cas-assisted SARS-CoV-2 detection assays are viewed as transformative solutions for POC diagnostic testing, but their lack of streamlined sample preparation and full integration within an automated and portable device hamper their potential for POC use. We report herein POC-CRISPR – a single-step CRISPR-Cas-assisted assay that incoporates sample preparation with minimal manual operation via facile magnetic-based nucleic acid concentration and transport. Moreover, POC-CRISPR has been adapted into a compact thermoplastic cartridge within a palm-sized yet fully-integrated and automated device. During analytical evaluation, POC-CRISPR was able detect 1 genome equivalent/μL SARS-CoV-2 RNA from a sample volume of 100 μL in < 30 min. When evaluated with 27 unprocessed clinical nasopharyngeal swab eluates that were pre-typed by standard RT-qPCR (C q values ranged from 18.3 to 30.2 for the positive samples), POC-CRISPR achieved 27 out of 27 concordance and could detect positive samples with high SARS-CoV-2 loads (C q < 25) in 20 min. • POC-CRISPR is a point-of-care amenable CRISPR-based sensing assay • The 1-step assay couples magnetofluidic sample preparation and CRISPR-aided RT-RPA • The assay is automated in a low-cost plastic cartridge and a palm-sized device • POC-CRISPR is evaluated with unprocessed clinical swab eluates (n = 27) • POC-CRISPR detects SARS-CoV-2 positive samples with high viral loads in 20 min [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF