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CoLAMP: CRISPR-based one-pot loop-mediated isothermal amplification enables at-home diagnosis of SARS-CoV-2 RNA with nearly eliminated contamination utilizing amplicons depletion strategy.
Cao, Yumeng; Lu, Xiao; Lin, Haosi; Rodriguez Serrano, Alan Fernando; Lui, Grace C Y; Hsing, I-Ming.
  • Cao Y; Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Hong Kong Special Administrative Region of China.
  • Lu X; Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Hong Kong Special Administrative Region of China.
  • Lin H; Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Hong Kong Special Administrative Region of China.
  • Rodriguez Serrano AF; Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Hong Kong Special Administrative Region of China.
  • Lui GCY; Department of Medicine & Therapeutics, The Chinese University of Hong Kong, Hong Kong Special Administrative Region of China.
  • Hsing IM; Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Hong Kong Special Administrative Region of China. Electronic address: kehsing@ust.hk.
Biosens Bioelectron ; 236: 115402, 2023 Sep 15.
Article in English | MEDLINE | ID: covidwho-2313386
ABSTRACT
Rapid point-of-care diagnostics, essential in settings such as airport on-site testing and home-based screening, displayed important implications for infectious disease control during the SARS-CoV-2 outbreak. However, the deployment of simple and sensitive assays in real-life scenarios still faces the concern of aerosol contamination. Here, we report an amplicon-depleting CRISPR-based one-pot loop-mediated isothermal amplification (CoLAMP) assay for point-of-care diagnosis of SARS-CoV-2 RNA. In this work, AapCas12b sgRNA is designed to recognize the activator sequence sited in the loop region of the LAMP product, which is crucial for exponential amplification. By destroying the aerosol-prone amplifiable products at the end of each amplification reaction, our design can significantly reduce the amplicons contamination that causes false positive results in point-of-care diagnostics. For at-home self-testing, we designed a low-cost sample-to-result device for fluorescence-based visual interpretation. As well, a commercial portable electrochemical platform was deployed as a proof-of-concept of ready-to-use point-of-care diagnostic systems. The field deployable CoLAMP assay can detect as low as 0.5 copies/µL of SARS-CoV-2 RNA in clinical nasopharyngeal swab samples within 40 min without the need for specialists for its operation.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Biosensing Techniques / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Biosens Bioelectron Journal subject: Biotechnology Year: 2023 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Biosensing Techniques / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Biosens Bioelectron Journal subject: Biotechnology Year: 2023 Document Type: Article