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Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin.
Guan, Bin; Frank, Karen M; Maldonado, José O; Beach, Margaret; Pelayo, Eileen; Warner, Blake M; Hufnagel, Robert B.
Afiliação
  • Guan B; Ophthalmic Genetics and Visual Function Branch, National Eye Institute, National Institutes of Health, 10 Center Drive, Building 10, Rm 10N109, Bethesda, MD 20892, USA.
  • Frank KM; Department of Laboratory Medicine, Clinical Center, Bethesda, MD 20892, USA.
  • Maldonado JO; AAV Biology Section, National Institute of Dental and Craniofacial Research, Bethesda, MD 20892, USA.
  • Beach M; Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA.
  • Pelayo E; Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA.
  • Warner BM; Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA.
  • Hufnagel RB; Salivary Disorders Unit, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA.
iScience ; 24(9): 102960, 2021 09 24.
Article em En | MEDLINE | ID: mdl-34396082
Current conventional detection of SARS-CoV-2 involves collection of a patient's sample with a nasopharyngeal swab, storage of the swab during transport in a viral transport medium, extraction of RNA, and quantitative reverse transcription PCR (RT-qPCR). We developed a simplified preparation method using a chelating resin, Chelex, which obviates RNA extraction during viral testing. Direct detection RT-qPCR and digital droplet PCR were compared to the current conventional method with RNA extraction for simulated samples and patient specimens. The heat treatment in the presence of Chelex markedly improved detection sensitivity as compared to heat alone, and lack of RNA extraction shortens the overall diagnostic workflow. Furthermore, the initial sample heating step inactivates SARS-CoV-2 infectivity, thus improving workflow safety. This fast RNA preparation and detection method is versatile for a variety of samples, safe for testing personnel, and suitable for standard clinical collection and testing on high-throughput platforms.
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Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2021 Tipo de documento: Article