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Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction.
Wei, Shan; Kohl, Esther; Djandji, Alexandre; Morgan, Stephanie; Whittier, Susan; Mansukhani, Mahesh; Hod, Eldad; D'Alton, Mary; Suh, Yousin; Williams, Zev.
  • Wei S; Department of Obstetrics and Gynecology, Columbia University Medical Center, New York, USA.
  • Kohl E; Department of Obstetrics and Gynecology, Columbia University Medical Center, New York, USA.
  • Djandji A; Department of Obstetrics and Gynecology, Columbia University Medical Center, New York, USA.
  • Morgan S; Department of Obstetrics and Gynecology, Columbia University Medical Center, New York, USA.
  • Whittier S; Department of Pathology and Cell Biology, Columbia University Medical Center, New York, USA.
  • Mansukhani M; Department of Pathology and Cell Biology, Columbia University Medical Center, New York, USA.
  • Hod E; Department of Pathology and Cell Biology, Columbia University Medical Center, New York, USA.
  • D'Alton M; Department of Obstetrics and Gynecology, Columbia University Medical Center, New York, USA.
  • Suh Y; Department of Obstetrics and Gynecology, Columbia University Medical Center, New York, USA.
  • Williams Z; Department of Genetics and Development, Columbia University Medical Center, New York, USA.
Sci Rep ; 11(1): 2402, 2021 01 28.
Artículo en Inglés | MEDLINE | ID: covidwho-1054048
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ABSTRACT
The COVID-19 pandemic has resulted in an urgent need for a rapid, point of care diagnostic testing that could be rapidly scaled on a worldwide level. We developed and tested a highly sensitive and robust assay based on reverse transcription loop mediated isothermal amplification (RT-LAMP) that uses readily available reagents and a simple heat block using contrived spike-in and actual clinical samples. RT-LAMP testing on RNA-spiked samples showed a limit of detection (LoD) of 2.5 copies/µl of viral transport media. RT-LAMP testing directly on clinical nasopharyngeal swab samples in viral transport media had an 85% positive percentage agreement (PPA) (17/20), and 100% negative percentage agreement (NPV) and delivered results in 30 min. Our optimized RT-LAMP based testing method is a scalable system that is sufficiently sensitive and robust to test for SARS-CoV-2 directly on clinical nasopharyngeal swab samples in viral transport media in 30 min at the point of care without the need for specialized or proprietary equipment or reagents. This cost-effective and efficient one-step testing method can be readily available for COVID-19 testing world-wide, especially in resource poor settings.
Asunto(s)

Texto completo: Disponible Colección: Bases de datos internacionales Base de datos: MEDLINE Asunto principal: ARN Viral / Técnicas de Amplificación de Ácido Nucleico / Técnicas de Diagnóstico Molecular / Prueba de COVID-19 / SARS-CoV-2 / COVID-19 Tipo de estudio: Estudios diagnósticos / Estudio pronóstico Límite: Humanos Idioma: Inglés Revista: Sci Rep Año: 2021 Tipo del documento: Artículo País de afiliación: S41598-021-81487-y

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Texto completo: Disponible Colección: Bases de datos internacionales Base de datos: MEDLINE Asunto principal: ARN Viral / Técnicas de Amplificación de Ácido Nucleico / Técnicas de Diagnóstico Molecular / Prueba de COVID-19 / SARS-CoV-2 / COVID-19 Tipo de estudio: Estudios diagnósticos / Estudio pronóstico Límite: Humanos Idioma: Inglés Revista: Sci Rep Año: 2021 Tipo del documento: Artículo País de afiliación: S41598-021-81487-y