High-Throughput CRISPR-Cas13 SARS-CoV-2 Test
| Autor: | Hannah J Thompson, Marisa C Tudino, Wahab A. Khan, Subha Singh, Heike Boisvert, Rachael E Barney, Elizabeth S Fiore, William J Blake, Gregory J. Tsongalis, Mary K Wilson, Jennifer M Peña, Brendan J Manning, Joel A Mowatt |
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| Rok vydání: | 2021 |
| Předmět: |
Diagnostic methods
Coronavirus disease 2019 (COVID-19) business.industry SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Concordance Biochemistry (medical) Clinical Biochemistry COVID-19 Molecular diagnostics Virology Sensitivity and Specificity High-Throughput Screening Assays COVID-19 Testing Nucleic acid CRISPR Medicine Humans RNA Viral CRISPR-Cas Systems business Throughput (business) |
| Zdroj: | Clinical chemistry. 68(1) |
| ISSN: | 1530-8561 |
| Popis: | Background The ability to control the spread of COVID-19 continues to be hampered by a lack of rapid, scalable, and easily deployable diagnostic solutions. Methods We developed a diagnostic method based on CRISPR (clustered regularly interspaced short palindromic repeats) that can deliver sensitive, specific, and high-throughput detection of Sudden Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2). The assay utilizes SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) for the qualitative detection of SARS-CoV-2 RNA and may be performed directly on a swab or saliva sample without nucleic acid extraction. The assay uses a 384-well format and provides results in Results Assay performance was evaluated with 105 (55 negative, 50 positive) remnant SARS-CoV-2 specimens previously tested using Food and Drug Administration emergency use authorized assays and retested with a modified version of the Centers for Disease Control and Prevention (CDC) quantitative PCR with reverse transcription (RT–qPCR) assay. When combined with magnetic bead-based extraction, the high-throughput SHERLOCK SARS-CoV-2 assay was 100% concordant (n = 60) with the CDC RT–qPCR. When used with direct sample addition the high-throughput assay was also 100% concordant with the CDC RT–qPCR direct method (n = 45). With direct saliva sample addition, the negative and positive percentage agreements were 100% (15/15, 95% CI: 81.8–100%) and 88% (15/17, 95% CI: 63.6–98.5%), respectively, compared with results from a collaborating clinical laboratory. Conclusions This high-throughput assay identifies SARS-CoV-2 from patient samples with or without nucleic acid extraction with high concordance to RT–qPCR methods. This test enables high complexity laboratories to rapidly increase their testing capacities with simple equipment. |
| Databáze: | OpenAIRE |
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