دورية أكاديمية
Real-Time, Multiplexed SHERLOCK for in Vitro Diagnostics.
| العنوان: | Real-Time, Multiplexed SHERLOCK for in Vitro Diagnostics. |
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| المؤلفون: | Pena JM; Sherlock Biosciences, Watertown, Massachusetts., Manning BJ; Sherlock Biosciences, Watertown, Massachusetts., Li X; Sherlock Biosciences, Watertown, Massachusetts., Fiore ES; Sherlock Biosciences, Watertown, Massachusetts., Carlson L; Sherlock Biosciences, Watertown, Massachusetts., Shytle K; Sherlock Biosciences, Watertown, Massachusetts., Nguyen PP; Sherlock Biosciences, Watertown, Massachusetts., Azmi I; Sherlock Biosciences, Watertown, Massachusetts., Larsen A; Sherlock Biosciences, Watertown, Massachusetts., Wilson MK; Sherlock Biosciences, Watertown, Massachusetts., Singh S; Sherlock Biosciences, Watertown, Massachusetts., DeMeo MC; Sherlock Biosciences, Watertown, Massachusetts., Ramesh P; Sherlock Biosciences, Watertown, Massachusetts., Boisvert H; Sherlock Biosciences, Watertown, Massachusetts. Electronic address: hboisvert@sherlock.bio., Blake WJ; Sherlock Biosciences, Watertown, Massachusetts. |
| المصدر: | The Journal of molecular diagnostics : JMD [J Mol Diagn] 2023 Jul; Vol. 25 (7), pp. 428-437. Date of Electronic Publication: 2023 Apr 23. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: United States NLM ID: 100893612 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1943-7811 (Electronic) Linking ISSN: 15251578 NLM ISO Abbreviation: J Mol Diagn Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2011- : New York : Elsevier Original Publication: Bethesda, MD : American Society for Investigative Pathology and the Association for Molecular Pathology, 1999- |
| مواضيع طبية MeSH: | COVID-19*/diagnosis , Clinical Laboratory Services*, Humans ; SARS-CoV-2/genetics ; Sensitivity and Specificity ; Molecular Diagnostic Techniques/methods ; COVID-19 Testing ; Nucleic Acid Amplification Techniques/methods |
| مستخلص: | The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has highlighted the need for simple, low-cost, and scalable diagnostics that can be widely deployed for rapid testing. Clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostics have emerged as a promising technology, but its implementation in clinical laboratories has been limited by the requirement of a separate amplification step prior to CRISPR-associated (Cas) enzyme-based detection. This article reports the discovery of two novel Cas12 enzymes (SLK9 and SLK5-2) that exhibit enzymatic activity at 60°C, which, when combined with loop-mediated isothermal amplification (LAMP), enable a real-time, single-step nucleic acid detection method [real-time SHERLOCK (real-time SLK)]. Real-time SLK was demonstrated to provide accurate results comparable to those from real-time quantitative RT-PCR in clinical samples, with 100% positive and 100% negative percent agreement. The method is further demonstrated to be compatible with direct testing (real-time SLK Direct) of samples from anterior nasal swabs, without the need for standard nucleic acid extraction. Lastly, SLK9 was combined with either Alicyclobacillus acidoterrestris AacCas12b or with SLK5-2 to generate a real-time, multiplexed CRISPR-based diagnostic assay for the simultaneous detection of SARS-CoV-2 and a human-based control in a single reaction, with sensitivity down to 5 copies/μL and a time to result of under 30 minutes. (Copyright © 2023 Association for Molecular Pathology and American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.) |
| تواريخ الأحداث: | Date Created: 20230423 Date Completed: 20230626 Latest Revision: 20240112 |
| رمز التحديث: | 20240112 |
| مُعرف محوري في PubMed: | PMC10122965 |
| DOI: | 10.1016/j.jmoldx.2023.03.009 |
| PMID: | 37088139 |
| قاعدة البيانات: | MEDLINE |
Full Text via ClinicalKey 
Full Text Finder | تدمد: | 1943-7811 |
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| DOI: | 10.1016/j.jmoldx.2023.03.009 |