Validation of one-step reverse transcription digital PCR assays for Norovirus GI.

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  • Author(s): Ko B;Ko B;Ko B; Shin T; Shin T; Kim B; Kim B; Lee DH; Lee DH
  • Source:
    Analytical biochemistry [Anal Biochem] 2024 Sep; Vol. 692, pp. 115576. Date of Electronic Publication: 2024 May 23.
  • Publication Type:
    Journal Article; Validation Study
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: United States NLM ID: 0370535 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1096-0309 (Electronic) Linking ISSN: 00032697 NLM ISO Abbreviation: Anal Biochem Subsets: MEDLINE
    • Publication Information:
      Publication: <2000- > : San Diego, CA : Elsevier
      Original Publication: Orlando Fl : Academic Press
    • Subject Terms:
      Norovirus*/genetics ; Norovirus*/isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction*/methods; RNA, Viral/genetics ; RNA, Viral/analysis ; Humans ; Caliciviridae Infections/diagnosis ; Caliciviridae Infections/virology
    • Abstract:
      Regular monitoring of Norovirus presence in environmental and food samples is crucial due to its high transmission rates and outbreak potential. For detecting Norovirus GI, reverse transcription qPCR method is commonly used, but its sensitivity can be affected by assay performance. This study shows significantly reduced assay performance in digital PCR or qPCR when using primers targeting Norovirus GI genome 5291-5319 (NC_001959), located on the hairpin of the predicted RNA structure. It is highly recommended to avoid this region in commercial kit development or diagnosis to minimizing potential risk of false negatives.
      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 The Authors. Published by Elsevier Inc. All rights reserved.)
    • Contributed Indexing:
      Keywords: Droplet based digital PCR; Nanoplate based digital PCR; Norovirus GI assay; One-step reverse transcription digital PCR; Reference material; Secondary structure
    • Accession Number:
      0 (RNA, Viral)
    • Publication Date:
      Date Created: 20240525 Date Completed: 20240614 Latest Revision: 20240614
    • Publication Date:
      20240615
    • Accession Number:
      10.1016/j.ab.2024.115576
    • Accession Number:
      38796118