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Detection of SARS-CoV-2 using non-commercial RT-LAMP regents and raw samples.
Alisa Alekseenko; Donal Barrett; Yerma Pareja-Sanchez; Rebecca Howard; Emilia Strandback; Henry Ampah-Korsah; Urška Rovšnik; Silvia Zuniga-Veliz; Alexander Klenov; Jayshna Malloo; Shenglong Ye; Xiyang Liu; Böjrn Reinius; Simon Elsässer; Tomas Nyman; Gustaf Sandh; Xiushan Yin; Vicent Pelechano.
Afiliación
  • Alisa Alekseenko; SciLifeLab, Department of Microbiology, Tumor and Cell Biology. Karolinska Institutet, Solna, Sweden
  • Donal Barrett; SciLifeLab, Department of Microbiology, Tumor and Cell Biology. Karolinska Institutet, Solna, Sweden
  • Yerma Pareja-Sanchez; SciLifeLab, Department of Microbiology, Tumor and Cell Biology. Karolinska Institutet, Solna, Sweden
  • Rebecca Howard; SciLifeLab, Department of Biochemistry and Biophysics, Stockholm University, 17121 Solna, Sweden
  • Emilia Strandback; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Solna, Sweden.
  • Henry Ampah-Korsah; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Solna, Sweden.
  • Urška Rovšnik; SciLifeLab, Department of Biochemistry and Biophysics, Stockholm University, 17121 Solna, Sweden
  • Silvia Zuniga-Veliz; Department of Clinical Microbiology, Karolinska University Hospital, Stockholm, Sweden.
  • Alexander Klenov; Hudak Lab, Department of Biology, York University, 4700 Keele St, Toronto, ON, M3J 1P3, Canada
  • Jayshna Malloo; Hudak Lab, Department of Biology, York University, 4700 Keele St, Toronto, ON, M3J 1P3, Canada
  • Shenglong Ye; Biotech & Biomedicine Science Shenyang Co. Ltd, Shenyang, 110000, China
  • Xiyang Liu; Biotech & Biomedicine Science Shenyang Co. Ltd, Shenyang, 110000, China
  • Böjrn Reinius; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Solna, Sweden.
  • Simon Elsässer; SciLifeLab, Department of Medical Biochemistry and Biophysics, Division of Genome Biology, Karolinska Institutet, Solna, Sweden.
  • Tomas Nyman; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Solna, Sweden.
  • Gustaf Sandh; Department of Clinical Microbiology, Karolinska University Hospital, Stockholm, Sweden.
  • Xiushan Yin; Applied Biology Laboratory, Shenyang University of Chemical Technology, 110142, Shenyang, China
  • Vicent Pelechano; SciLifeLab, Department of Microbiology, Tumor and Cell Biology. Karolinska Institutet, Solna, Sweden.
Preprint en En | PREPRINT-MEDRXIV | ID: ppmedrxiv-20179507
ABSTRACT
Abstract RT-LAMP detection of SARS-CoV-2 has been shown as a valuable approach to scale up COVID-19 diagnostics and thus contribute to limiting the spread of the disease. Here we present the optimization of highly cost-effective in-house produced enzymes, and we benchmark their performance against commercial alternatives. We explore the compatibility between multiple DNA polymerases with high strand-displacement activity and thermostable reverse transcriptases required for RT-LAMP. We optimize reaction conditions and demonstrate their applicability using both synthetic RNA and clinical patient samples. Finally, we validated the optimized RT-LAMP assay for the detection of SARS-CoV-2 in raw nasopharyngeal samples from 184 patients. We anticipate that optimized and affordable reagents for RT-LAMP will facilitate the expansion of SARS-CoV-2 testing globally, especially in sites and settings with limited economic resources.
Licencia
cc_by_nc_nd
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Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-MEDRXIV Tipo de estudio: Prognostic_studies Idioma: En Año: 2020 Tipo del documento: Preprint
Texto completo
Añadir a Mi BVS
Imprimir
XML
Buscar en Google
Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-MEDRXIV Tipo de estudio: Prognostic_studies Idioma: En Año: 2020 Tipo del documento: Preprint