RT Journal Article
SR Electronic
T1 Real-time colorimetric LAMP methodology for quantitative nucleic acids detection at the point-of-care
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.07.22.215251
DO 10.1101/2020.07.22.215251
A1 Papadakis, George
A1 Pantazis, Alexandros K.
A1 Fikas, Nikolaos
A1 Chatziioannidou, Stella
A1 Michaelidou, Kleita
A1 Pogka, Vasiliki
A1 Megariti, Maria
A1 Vardaki, Maria
A1 Giarentis, Konstantinos
A1 Heaney, Judith
A1 Nastouli, Eleni
A1 Karamitros, Timokratis
A1 Mentis, Andreas
A1 Agelaki, Sofia
A1 Gizeli, Electra
YR 2020
UL http://biorxiv.org/content/early/2020/07/24/2020.07.22.215251.abstract
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