LateralflowAssay for Disposable nonInvasive Easy-to-use Smartphone-based biosensor for ovarian Cancer diagnosis....- Finanziamento dell’Unione Europea – NextGenerationEU – missione 4, componente 2, investimento 1.1.

Conventional methods for the diagnosis of several cancers, including ovarian cancer (OC), are based on invasive and painful methods that are not suitable for frequent screening, which is, however, crucial for early diagnosis and for increasing the chance of survival. Thus, the development of sensitive, portable, low-cost and easy-to-use (bio)sensors for non-invasively detecting specific biomarkers in biological fluids are urgently needed for enabling frequent screening of a large part of the population to early detect the tumor and timely intervene. MicroRNA (miRNAs) are strong candidates as diagnostic markers for cancers since their expression profile significantly changes even at the early stage of the disease. As an example, several specific miRNAs are significantly abnormally expressed in OC and represent ideal blood-based diagnostic biomarkers. Furthermore, the diagnostic significance of measuring levels of miRNA is strongly increased when single information is combined. The project aims at the development of a portable and low-cost biosensor for the simultaneous quantification of different miRNAs specific for OC in capillary blood, combining the simple and easy-to-use nucleic acid lateral flow assay (NALFA) with optical based detection (visual-colorimetric and chemiluminescent (CL)) and exploiting smartphone-based signal measurement and processing. The new biosensor will be based on the LFA technique that consists in the use of a nitrocellulose porous membrane in which probes specific for the recognition of target analytes are immobilized in spatially confined zones (test zones). While few microliters of blood sample are flowing through the strip, each target miRNA will be captured by a specific DNA capture probe and then detected upon hybridization with a second DNA probe suitably labeled for optical-based detection. The colorimetric or CL signals will be detected using a smartphone’s camera and digital images will be elaborated by using an ad-hoc Application to reach quantitative measuring. The device will comprise a cartridge hosting the LFA strip and the reagents necessary for the CL assay, and a smartphone adaptor creating a “mini dark-box” and ensuring correct positioning of the strip in front of the camera. All the accessories will be produced exploiting state-of-the-art 3D printing technology. The developed App will be used also to combine quantitative information about target analytes, providing a score, which could represent an alarm for directing to confirmation diagnostics. The development of minimally invasive tests for OC early diagnosis could be a breakthrough in the fight against this malignancy, enabling large-scale screening at low cost and could represent a prototype for the further development of a new generation of multiplexing miRNA biosensors, exploiting the same strategy for the effective, early and sensitive diagnosis of other tumors.