Este articulo es un Preprint
Los preprints son informes de investigación preliminares que no han sido certificados por revisión por pares. No deben considerarse para guiar la práctica clínica o los comportamientos relacionados con la salud y no deben publicarse en los medios como información establecida.
Los preprints publicados en línea permiten a los autores recibir comentarios rápidamente, y toda la comunidad científica puede evaluar de forma independiente el trabajo y responder adecuadamente. Estos comentarios se publican junto con los preprints para que cualquiera pueda leer y servir como una revisión pospublicación.
Screening for SARS-CoV-2 infections with colorimetric RT-LAMP and LAMP sequencing
Preprint
en En
| PREPRINT-MEDRXIV
| ID: ppmedrxiv-20092288
ABSTRACT
The COVID-19 pandemic caused by the novel SARS-CoV-2 virus poses a significant public-health problem. In order to control the pandemic, rapid tests for detecting existing infections and assessing virus spread are critical. Approaches to detect viral RNA based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) hold outstanding promise towards greatly simplified and broadly applicable testing methods. RT-LAMP assays appear more robust than qPCR-based methods and only require incubation at a constant temperature, thus eliminating the need for sophisticated instrumentation. Here, we tested a two-color RT-LAMP protocol using clinical SARS-CoV-2 samples and also established a protocol that does not require prior RNA isolation ("swab-to-RT-LAMP"). Our study is based on several hundred clinical patient samples with a wide range of viral loads, thus allowing, for the first time, to accurately determine the sensitivity and specificity of the RT-LAMP assay for the detection of SARS-CoV-2 in patients. We found that RT-LAMP can reliably detect SARS-CoV-2 samples with a qPCR threshold cycle number (CT value) of up to 30 in the standard RT-qPCR assay. We used both, either purified RNA or direct pharyngeal swab specimens and showed that RT-LAMP assays have, despite a decreased sensitivity compared to RT-qPCR, excellent specificity. We also developed a multiplexed LAMP-sequencing protocol as a diagnostic and validation procedure to detect and record the outcome of RT-LAMP assays. LAMP-sequencing is fully scalable and can assess the results of thousands of LAMP reactions in parallel. Finally, we propose applications of RT-LAMP based assays for SARS-CoV-2 detection.
cc_by_nd
Texto completo:
1
Colección:
09-preprints
Base de datos:
PREPRINT-MEDRXIV
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Idioma:
En
Año:
2020
Tipo del documento:
Preprint