Research project

FreeATPOC -Towards an instrument-free future of molecular diagnostics at the point-of-care (DBA.AD001.401)

Thematic area

Biology, agriculture and food sciences

Project area

Intensificazione sostenibile delle produzioni agroalimentari e forestali (DBA.AD001)

Structure responsible for the research project

Institute for Sustainable Plant Protection (IPSP)

Project manager

ANGELANTONIO MINAFRA
Phone number: 0805443065
Email: ANGELANTONIO.MINAFRA@IPSP.CNR.IT

Abstract

Emerging technologies for application at the point of care have started gaining the molecular diagnostics market. available methods are divided in two groups: sophisticated instruments for ultra-sensitive temperature-regulated enzymatic amplification of nucleic acids and extremely simple paper- based immuno-diagnostic kits. Our vision is an instrument-free approach that combines the advantages of both groups, i.e., takes advantage of enzymatically-amplified nucleic acids detection. novel diagnostic tools for genetic amplification based on ligases, polymerases and restriction enzymes operating at ambient temperature will be developed, combined with quantitative smartphone colorimetric/UV detection. The global applicability of the new approach will be demonstrated during infectious disease, testing in human samples and plant pathogens (Xylella fastidiosa) in plant- tissues. In all cases, we aim for a time-to-result of less than 60 min, demonstrated sensitivities down to the clinically/field relevant values and a final cost in the order of $1 per assay.

Goals

This new generation of instrument-free molecular diagnostics is expected to revolutionize nucleic acid analysis at the POC but also in applications beyond resulting in substantial societal as well as economic benefits.
1. Novel molecular diagnostic tools: Development of assays for DNA/RNA digestion and amplification using a reaction cartridge and at RT based on restriction enzymes, polymerases and ligases of high specificity and single base discrimination capability; work directly in crude samples without any or with minimal pre-treatment.
2. Quantitative nucleic acid detection via smartphone: Combination of the above assays with a pH-sensitive colorimetric dye coupled to the production of amplicons; quantitative analysis via any smartphone (end point) or an on-line (real-time) camera through a purpose-developed software and smartphone application (not specific to smartphone features).
3. Demonstrate global applicability: Provide a proof-of-concept during the detection of infectious diseases in the field (Xylella), demonstrating the required limits of detection (1-1000 virus particles or bacterial genome copies)

Start date of activity

01/01/2020

Keywords

molecular detection, Xylella, olive

Last update: 08/02/2023