Multifunctional self-priming hairpin probe-based isothermal nucleic acid amplification and its applications for COVID-19 diagnosis.

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  • Additional Information
    • Source:
      Publisher: Elsevier Advanced Technology Country of Publication: England NLM ID: 9001289 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4235 (Electronic) Linking ISSN: 09565663 NLM ISO Abbreviation: Biosens Bioelectron Subsets: MEDLINE
    • Publication Information:
      Publication: Oxford : Elsevier Advanced Technology
      Original Publication: [Barking, Essex, England] : Elsevier Applied Science, 1989-
    • Subject Terms:
      Nucleic Acids* ; COVID-19*/diagnosis ; Biosensing Techniques*/methods; Humans ; COVID-19 Testing ; Nucleic Acid Amplification Techniques/methods ; SARS-CoV-2/genetics ; Sensitivity and Specificity
    • Abstract:
      We herein present a multifunctional self-priming hairpin probe-based isothermal amplification, termed MSH, enabling one-pot detection of target nucleic acids. The sophisticatedly designed multifunctional self-priming hairpin (MSH) probe recognizes the target and rearranges to prime itself, triggering the amplification reaction powered by the continuously repeated extension, nicking, and target recycling. As a consequence, a large number of double-stranded DNA (dsDNA) amplicons are produced that could be monitored in real-time using a dsDNA-intercalating dye. Based on this unique design approach, the nucleocapsid (N) and the open reading frame 1 ab (ORF1ab) genes of SARS-CoV-2 were successfully detected down to 1.664 fM and 0.770 fM, respectively. The practical applicability of our method was validated by accurately diagnosing 60 clinical samples with 93.33% sensitivity and 96.67% specificity. This isothermal one-pot MSH technique holds great promise as a point-of-care testing protocol for the reliable detection of a wide spectrum of pathogens, particularly in resource-limited settings.
      Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
      (Copyright © 2024 Elsevier B.V. All rights reserved.)
    • Contributed Indexing:
      Keywords: COVID-19; Isothermal amplification; Molecular diagnostics; SARS-CoV-2; Self-priming hairpin probe
    • Accession Number:
      0 (Nucleic Acids)
    • Publication Date:
      Date Created: 20240307 Date Completed: 20240319 Latest Revision: 20240319
    • Publication Date:
      20240319
    • Accession Number:
      10.1016/j.bios.2024.116147
    • Accession Number:
      38452568