Microfluidic cartridge with integrated array of amorphous silicon photosensors for chemiluminescence detection of viral DNA

Portable and simple analytical devices based on microfluidics with chemiluminescence detection are particularly attractive for point-of-care applications, offering high detectability and specificity in a simple and miniaturized analytical format. Particularly relevant for infectious disease diagnosis is the ability to sensitively and specifically detect target nucleic acid sequences in biological fluids. To reach the goal of real-life applications for such devices, however, several technological challenges related to full device integration are still to be solved, one key aspect regarding on-chip integration of the chemiluminescence signal detection device. Nowadays, the most promising approach is on-chip integration of thin-film photosensors. We recently proposed a portable cartridge with microwells aligned with an array of hydrogenated amorphous silicon (a-Si:H) photosensors, reaching attomole level limits of detection for different chemiluminescence model reactions. Herein, we explore its applicability and performance for multiplex and quantitative detection of viral DNA. In particular, the cartridge was modified to accommodate microfluidic channels and, upon immobilization of three oligonucleotide probes in different positions along each channel, each specific for a genotype of Parvovirus B19, viral nucleic acid sequences were captured and detected. With this system, taking advantage of oligoprobes specificity, chemiluminescence detectability, and photosensor sensitivity, accurate quantification of target analytes down to 70 pmol L-1 was obtained for each B19 DNA genotype, with high specificity and multiplexing ability. Results confirm the good detection capabilities and assay applicability of the proposed system, prompting the development of innovative portable analytical devices with enhanced sensitivity and multiplexed capabilities.