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Real-time colorimetric LAMP methodology for quantitative nucleic acids detection at the point-of-care

By George Papadakis, Alexandros K. Pantazis, Nikolaos Fikas, Stella Chatziioannidou, Kleita Michaelidou, Vasiliki Pogka, Maria Megariti, Maria Vardaki, Konstantinos Giarentis, Judith Heaney, Eleni Nastouli, Timokratis Karamitros, Andreas Mentis, Sofia Agelaki, Electra Gizeli

Posted 24 Jul 2020
bioRxiv DOI: 10.1101/2020.07.22.215251

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 Statement G.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

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