Surface Functionalized Anodic Aluminum Oxide Membrane for Opto-Nanofluidic SARS-CoV-2 Genomic Target Detection

An ultra-thin and highly sensitive SARS-CoV-2 detection platform was demonstrated using a nano-porous anodic aluminum oxide (AAO) membrane. The membrane surface was functionalized to enable efficient trapping and identification of SARS-CoV-2 genomic targets through DNA-DNA and DNA-RNA hybridization. To immobilize the probe oligonucleotides on the AAO membrane, the pore surface was first coated with the linking reagents, 3-aminopropyltrimethoxysilane (APTMS) and glutaraldehyde (GA), by a compact vacuum infiltration module. After that, complementary target oligos with fluorescent modifier was pulled and infiltrated into the nano-fluidic channels formed by the AAO pores. The fluorescent signal applying the AAO membrane sensors was two orders stronger than a flat glass template. In addition, the dependence between the nano-pore size and the fluorescent intensity was evaluated. The optimized pore diameter d is 200 nm, which can accommodate the assembled oligonucleotide and aminosilane layers without blocking the AAO nano-fluidic channels. Our DNA functionalized membrane sensor is an accurate and high throughput platform supporting rapid virus tests, which is critical for population-wide diagnostic applications result in a page being rejected by search engines.