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Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples.
Alekseenko, Alisa; Barrett, Donal; Pareja-Sanchez, Yerma; Howard, Rebecca J; Strandback, Emilia; Ampah-Korsah, Henry; Rovsnik, Urska; Zuniga-Veliz, Silvia; Klenov, Alexander; Malloo, Jayshna; Ye, Shenglong; Liu, Xiyang; Reinius, Björn; Elsässer, Simon J; Nyman, Tomas; Sandh, Gustaf; Yin, Xiushan; Pelechano, Vicent.
Afiliación
  • Alekseenko A; SciLifeLab, Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Tomtebodavägen 23A, 17165, Solna, Sweden.
  • Barrett D; SciLifeLab, Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Tomtebodavägen 23A, 17165, Solna, Sweden.
  • Pareja-Sanchez Y; SciLifeLab, Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Tomtebodavägen 23A, 17165, Solna, Sweden.
  • Howard RJ; SciLifeLab, Department of Biochemistry and Biophysics, Stockholm University, 17121, Solna, Sweden.
  • Strandback E; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Solna, Sweden.
  • Ampah-Korsah H; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Solna, Sweden.
  • Rovsnik U; SciLifeLab, Department of Biochemistry and Biophysics, Stockholm University, 17121, Solna, Sweden.
  • Zuniga-Veliz S; Department of Clinical Microbiology, Karolinska University Hospital, Stockholm, Sweden.
  • Klenov A; Hudak Lab, Department of Biology, York University, 4700 Keele St, Toronto, ON, M3J 1P3, Canada.
  • Malloo J; Hudak Lab, Department of Biology, York University, 4700 Keele St, Toronto, ON, M3J 1P3, Canada.
  • Ye S; Biotech and Biomedicine Science (Shenyang) Co. Ltd, Shenyang, 110000, China.
  • Liu X; Biotech and Biomedicine Science (Shenyang) Co. Ltd, Shenyang, 110000, China.
  • Reinius B; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Solna, Sweden.
  • Elsässer SJ; SciLifeLab, Department of Medical Biochemistry and Biophysics, Division of Genome Biology, Karolinska Institutet, Solna, Sweden.
  • Nyman T; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Solna, Sweden.
  • Sandh G; Department of Clinical Microbiology, Karolinska University Hospital, Stockholm, Sweden.
  • Yin X; SciLifeLab, Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Tomtebodavägen 23A, 17165, Solna, Sweden.
  • Pelechano V; Biotech and Biomedicine Science (Shenyang) Co. Ltd, Shenyang, 110000, China.
Sci Rep ; 11(1): 1820, 2021 01 19.
Article en En | MEDLINE | ID: mdl-33469065
RT-LAMP detection of SARS-CoV-2 has been shown to be 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 validate the optimized RT-LAMP assay for the detection of SARS-CoV-2 in unextracted heat-inactivated 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 where the need for large scale testing cannot be met by commercial alternatives.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Amplificación de Ácido Nucleico / Técnicas de Diagnóstico Molecular / SARS-CoV-2 / COVID-19 Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Sci Rep Año: 2021 Tipo del documento: Article País de afiliación: Suecia Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Amplificación de Ácido Nucleico / Técnicas de Diagnóstico Molecular / SARS-CoV-2 / COVID-19 Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Sci Rep Año: 2021 Tipo del documento: Article País de afiliación: Suecia Pais de publicación: Reino Unido