Dataset Information

Ultra-sensitive and high-throughput CRISPR-p owered COVID-19 diagnosis.

ABSTRACT: Recent research suggests that SARS-CoV-2-infected individuals can be highly infectious while asymptomatic or pre-symptomatic, and that an infected person may infect 5.6 other individuals on average. This situation highlights the need for rapid, sensitive SARS-CoV-2 diagnostic assays capable of high-throughput operation that can preferably utilize existing equipment to facilitate broad, large-scale screening efforts. We have developed a CRISPR-based assay that can meet all these criteria. This assay utilizes a custom CRISPR Cas12a/gRNA complex and a fluorescent probe to detect target amplicons produced by standard RT-PCR or isothermal recombinase polymerase amplification (RPA), to allow sensitive detection at sites not equipped with real-time PCR systems required for qPCR diagnostics. We found this approach allowed sensitive and robust detection of SARS-CoV-2 positive samples, with a sample-to-answer time of ~50 min, and a limit of detection of 2 copies per sample. CRISPR assay diagnostic results obtained nasal swab samples of individuals with suspected COVID-19 cases were comparable to paired results from a CDC-approved quantitative RT-PCR (RT-qPCR) assay performed in a state testing lab, and superior to those produced by same assay in a clinical lab, where the RT-qPCR assay exhibited multiple invalid or inconclusive results. Our assay also demonstrated greater analytical sensitivity and more robust diagnostic performance than other recently reported CRISPR-based assays. Based on these findings, we believe that a CRISPR-based fluorescent application has potential to improve current COVID-19 screening efforts.

SUBMITTER: Huang Z

PROVIDER: S-EPMC7245202 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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Ultra-sensitive and high-throughput CRISPR-p owered COVID-19 diagnosis.

Huang Zhen Z Tian Di D Liu Yang Y Lin Zhen Z Lyon Christopher J CJ Lai Weihua W Fusco Dahlene D Drouin Arnaud A Yin Xiaoming X Hu Tony T Ning Bo B

Biosensors & bioelectronics 20200523

Recent research suggests that SARS-CoV-2-infected individuals can be highly infectious while asymptomatic or pre-symptomatic, and that an infected person may infect 5.6 other individuals on average. This situation highlights the need for rapid, sensitive SARS-CoV-2 diagnostic assays capable of high-throughput operation that can preferably utilize existing equipment to facilitate broad, large-scale screening efforts. We have developed a CRISPR-based assay that can meet all these criteria. This as ...[more]

PMID: 32553350

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Project description:The urgent need for diagnostics and therapeutics against the COVID-19 pandemic has shown the great potential of antibodies, proteomics and metabolomics in this direction. Several clinical trials are underway using antibodies from COVID-19 patients that show very specific and strong binding to viral proteins leading to neutralization. On the other hand, proteomic and metabolomic profiles of COVID-19 patients present novel diagnostic biomarkers to predict patient outcomes and enable the development of personalized therapeutics to target the dysregulated pathways, as revealed by those profiles. Here, we discuss how studies based on antibodies, proteomics and metabolomics contribute to the development of diagnostics and therapeutics against COVID-19. The elegant technology can extend to high-throughput, rapid and reliable drug discovery strategies of the future. Lay abstract In order to prevent and treat the ongoing COVID-19 pandemic, several groups around the world are focused on a detailed understanding of the biology of SARS-CoV-2 infection, the biological events occurring inside the patients and the response of the patients to the infection. SARS-CoV-2 is the coronavirus that causes COVID-19. Some of the approaches to combat the pandemic have provided important results that can help toward therapeutic developments against COVID-19. This review discusses three such areas – antibody treatment to prevent COVID-19 infection, analysis of changes in protein profiles of COVID-19 patients, and analysis of metabolism or energy-related changes in COVID-19 patients. Antibodies are molecules produced in the host’s body as a defense response to infection. Antibodies extracted from patients who recovered from COVID-19 have been used to successfully manufacture large amounts of antibodies to treat COVID-19 patients. The analysis of protein and metabolism profiles of COVID-19 patients has shown that several proteins and metabolism-related entities in the body are either upregulated or downregulated in COVID-19. These abnormal levels can either be attenuated by medical intervention or can be monitored as indicators of COVID-19 diagnosis. Overall, antibodies, protein and metabolism profiles are three important tools, among several others, that are helpful in combating the COVID-19 pandemic.

Dataset Information

Ultra-sensitive and high-throughput CRISPR-p owered COVID-19 diagnosis.

Publications

Ultra-sensitive and high-throughput CRISPR-p owered COVID-19 diagnosis.

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