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Development and application of sensitive, specific, and rapid CRISPR-Cas13-based diagnosis.
Shihong Gao, David; Zhu, Xiaodong; Lu, Binfeng.
  • Shihong Gao D; Department of Immunology, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
  • Zhu X; Department of Biological Sciences, Dietrich School of Arts and Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
  • Lu B; Department of Immunology, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
J Med Virol ; 93(7): 4198-4204, 2021 Jul.
Article in English | MEDLINE | ID: covidwho-1263091
ABSTRACT
Nucleic acid detection is a necessary part of medical treatment and fieldwork. However, the current detection technologies are far from ideal. A lack of timely and accessible testing for identifying cases and close contacts has allowed severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causative virus of the ongoing coronavirus disease-2019 (COVID-19) pandemic, to spread uncontrollably. The slow and expensive detection of mutations-predictors for chronic diseases such as cancer-form a barrier to personalized treatment. A recently developed diagnostic assay is ideal and field-ready-it relies on CRISPR-Cas13. CRISPR-Cas13 works similarly to other CRISPR systems Cas13 is guided by a crRNA to cleave next to a specific RNA target sequence. Additionally, Cas13 boasts a unique collateral cleavage activity; collateral cleavage of a fluorescent reporter detects the presence of the target sequence in sample RNA. This system forms the basis of CRISPR-Cas13 diagnostic assays. CRISPR-Cas13 assays have >95% sensitivity and >99% specificity. Detection is rapid (<2 h), inexpensive ($0.05 per test), and portable-a test using lateral flow strips is akin to a pregnancy test. The recent adaptation of micro-well chips facilitates high-level multiplexing and is high-throughput. In this review, we cover the development of CRISPR-Cas13 assays for medical diagnosis, discuss the advantages of CRISPR-Cas13-based diagnosis over the traditional reverse transcription polymerase chain reaction (RT-PCR), and present examples of detection from real patient samples.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Clustered Regularly Interspaced Short Palindromic Repeats / CRISPR-Cas Systems / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study / Qualitative research Limits: Humans Language: English Journal: J Med Virol Year: 2021 Document Type: Article Affiliation country: Jmv.26889

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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Clustered Regularly Interspaced Short Palindromic Repeats / CRISPR-Cas Systems / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study / Qualitative research Limits: Humans Language: English Journal: J Med Virol Year: 2021 Document Type: Article Affiliation country: Jmv.26889