Fluorescent Label-Free Aptasensor Integrated in a Lab-on-Chip System for the Detection of Ochratoxin A in Beer and Wheat.

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.