This paper reports on the development of a fluorescent label-free aptamer assay integrated in a lab-on-chip (LoC) system for the detection of Ochratoxin A (OTA). The detection system relies on the integration, on a single glass substrate, of an array of amorphous silicon photosensors and a long pass interferential filter. The aptamer assay, integrated into the microfluidic network, is an aptasensor having affinity versus OTA, selected as a case study. The fluorescent molecule is a "light switch" complex [Ru(phen)2(dppz)]2+. The aptamer is directly anchored into a layer of poly(2-hydroxyethyl methacrylate) polymer brushes grown inside the channels. The fluorophore is intercalated between the base pairs of the aptamer. Upon the interaction of OTA with the aptasensor, a change of the aptamer conformation causes the release of the fluorophore, yielding a decrease of the fluorescent signal detected by the array of the amorphous silicon photosensors positioned underneath the microfluidic network. The developed LoC is a portable system capable of performing the analysis with a small volume of sample (about 10 μL) in a short time (5 min) with a limit of detection for OTA equal to 1.3 ng/mL. The LoC has been applied for the detection of OTA (5-200 ng/mL) in beer and wheat samples.

Fluorescent label-free aptasensor integrated in a lab-on-chip system for the detection of Ochratoxin A in beer and wheat / Costantini, F.; Lovecchio, N.; Ruggi, A.; Manetti, C.; Nascetti, A.; Reverberi, M.; De Cesare, G.; Caputo, D.. - In: ACS APPLIED BIO MATERIALS. - ISSN 2576-6422. - 2:12(2019), pp. 5880-5887. [10.1021/acsabm.9b00831]

Fluorescent label-free aptasensor integrated in a lab-on-chip system for the detection of Ochratoxin A in beer and wheat

Costantini F.
;
Lovecchio N.;Manetti C.;Nascetti A.;Reverberi M.;De Cesare G.;Caputo D.
2019

Abstract

This paper reports on the development of a fluorescent label-free aptamer assay integrated in a lab-on-chip (LoC) system for the detection of Ochratoxin A (OTA). The detection system relies on the integration, on a single glass substrate, of an array of amorphous silicon photosensors and a long pass interferential filter. The aptamer assay, integrated into the microfluidic network, is an aptasensor having affinity versus OTA, selected as a case study. The fluorescent molecule is a "light switch" complex [Ru(phen)2(dppz)]2+. The aptamer is directly anchored into a layer of poly(2-hydroxyethyl methacrylate) polymer brushes grown inside the channels. The fluorophore is intercalated between the base pairs of the aptamer. Upon the interaction of OTA with the aptasensor, a change of the aptamer conformation causes the release of the fluorophore, yielding a decrease of the fluorescent signal detected by the array of the amorphous silicon photosensors positioned underneath the microfluidic network. The developed LoC is a portable system capable of performing the analysis with a small volume of sample (about 10 μL) in a short time (5 min) with a limit of detection for OTA equal to 1.3 ng/mL. The LoC has been applied for the detection of OTA (5-200 ng/mL) in beer and wheat samples.
2019
amorphous silicon; aptamer; assay; fluorescence; microfluidics; thin-film technology
01 Pubblicazione su rivista::01a Articolo in rivista
Fluorescent label-free aptasensor integrated in a lab-on-chip system for the detection of Ochratoxin A in beer and wheat / Costantini, F.; Lovecchio, N.; Ruggi, A.; Manetti, C.; Nascetti, A.; Reverberi, M.; De Cesare, G.; Caputo, D.. - In: ACS APPLIED BIO MATERIALS. - ISSN 2576-6422. - 2:12(2019), pp. 5880-5887. [10.1021/acsabm.9b00831]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1366819
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