PT  - JOURNAL ARTICLE
AU  - Papadakis, George
AU  - Pantazis, Alexandros K.
AU  - Fikas, Nikolaos
AU  - Chatziioannidou, Stella
AU  - Michaelidou, Kleita
AU  - Pogka, Vasiliki
AU  - Megariti, Maria
AU  - Vardaki, Maria
AU  - Giarentis, Konstantinos
AU  - Heaney, Judith
AU  - Nastouli, Eleni
AU  - Karamitros, Timokratis
AU  - Mentis, Andreas
AU  - Agelaki, Sofia
AU  - Gizeli, Electra
TI  - Real-time colorimetric LAMP methodology for quantitative nucleic acids detection at the point-of-care
AID  - 10.1101/2020.07.22.215251
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.07.22.215251
4099  - http://biorxiv.org/content/early/2020/07/24/2020.07.22.215251.short
4100  - http://biorxiv.org/content/early/2020/07/24/2020.07.22.215251.full
AB  - Most methods applied to nucleic acids’ detection at the point-of-care require either expensive and mostly bench-top instruments or simpler inexpensive systems providing qualitative results. Truly decentralized approaches to reliable, quantitative and affordable diagnostics are still missing. Here, we report the development of real-time quantitative colorimetric LAMP based on a portable and cost-effective device, the use of which requires minimal training. Main advantages of the method are the rapid analysis time (<30min); quantification over a large dynamic range (9 log units); ability to work with crude samples (saliva, tissue); demonstrated low detection limit (1-10 copies); smartphone-operation and fast prototyping (3D-printing). The system’s broad detection capability is demonstrated during infectious diseases-testing for COVID-19 and pharmacogenetics for BRAF V600E mutation testing. Validation studies showed 97.4% and 100% agreement with qRT-PCR for SARS-CoV-2 RNA detection extracted from positive and negative patients’ samples (89), respectively; and 100% agreement with ddPCR and Sanger sequencing for BRAF V600E mutation detection from 12 clinical biopsy samples. The new methodology provides a needed solution for affordable healthcare at the point-of-care, with emphasis on global diagnostics.Competing Interest StatementG.P., A.K.P., N.F. and E.G. are the co-founders of BIOPIX DNA TECHNOLOGY P.C. The other authors declare no competing interests