High-Throughput CRISPR-Cas13 SARS-CoV-2 Test.
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 sensitiv...
| Publicado en: | Clinical Chemistry Vol. 68; no. 1; pp. 172 - 181 |
|---|---|
| Autores principales: | , , , , , , , , , , , , |
| Formato: | pictorial research tables/charts Journal Article |
| Publicado: |
Oxford University Press / USA
Jan2022
|
| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=154525753&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 154525753 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 00099147 10CS jtl: Clinical Chemistry issn: 00099147 maglogo: N pubinfo: dt: Jan2022 vid: 68 iid: 1 pid: 622 pub: Oxford University Press / USA artinfo: ui: 154525753 154525753 154525753 10.1093/clinchem/hvab238 154525753 ppf: 172 ppct: 9 formats: fmt: @attributes: type: P tig: atl: High-Throughput CRISPR-Cas13 SARS-CoV-2 Test. aug: au: Manning, Brendan J. Khan, Wahab A. Peña, Jennifer M. Fiore, Elizabeth S. Boisvert, Heike Tudino, Marisa C. Barney, Rachael E. Wilson, Mary K. Singh, Subha Mowatt, Joel A. Thompson, Hannah J. Tsongalis, Gregory J. Blake, William J. affil: R&D Department, Sherlock Biosciences, Boston, MA sug: subj: COVID-19 Diagnosis COVID-19 Testing Methods Clustered Regularly Interspaced Short Palindromic Repeats Analysis Molecular Diagnostic Techniques Human Reverse Transcriptase Polymerase Chain Reaction Comparative Studies Descriptive Statistics Saliva Confidence Intervals Sensitivity and Specificity Enzyme Tests ab: 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 <1 hour. 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. pubtype: Academic Journal doctype: pictorial research tables/charts Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
|---|