Anti-DNA:RNA Antibodies and Silicon Photonic Microring Resonators: Increased Sensitivity for Multiplexed microRNA Detection

Abstract

In this paper, we present a method for the ultrasensitive detection of microRNAs (miRNAs) utilizing an antibody that specifically recognizes DNA:RNA heteroduplexes, using a silicon photonic microring resonator array transduction platform. Microring resonator arrays are covalently functionalized with DNA capture probes that are complementary to solution phase miRNA targets. Following hybridization on the sensor, the anti-DNA:RNA antibody is introduced and binds selectively to the heteroduplexes, giving a larger signal than the original miRNA hybridization due to the increased mass of the antibody, as compared to the 22 oligoribonucleotide. Furthermore, the secondary recognition step is performed in neat buffer solution and at relatively higher antibody concentrations, facilitating the detection of miRNAs of interest. The intrinsic sensitivity of the microring resonator platform coupled with the amplification provided by the anti-DNA:RNA antibodies allows for the detection of microRNAs at concentrations as low as 10 pM (350 attomoles). The simplicity and sequence generality of this amplification method position it as a promising tool for high-throughput, multiplexed miRNA analysis, as well as a range of other RNA based detection applications.

An electrochemical ELISA-like immunosensor for miRNAs detection based on screen-printed gold electrodes modified with reduced graphene oxide and carbon nanotubes

“…The breakthrough consists in using antibodies specifically directed to RNA.DNA hybrids, which can thereafter be exploited for RNA detection through classical Enzyme Linked ImmunoSorbent Assay (ELISA). However, there are very few recent research works mentioning such miRNA detection through immunoassays (Qavi et al, 2011;Dong et al, 2012c;Sípová et al, 2010). In these studies, antibodies bind miRNA.DNA hybrids attached on a surface and, due to their steric hindrance, enhanced the signal for surface plasmon resonance (SPR), silicon microring photonic resonators detection or electrochemical detection (Tran et al, 2013b).…”

High sensitivity and label-free oligonucleotides detection using photonic bandgap sensing structures biofunctionalized with molecular beacon probes

“…Among the different transduction technologies considered for these developments, planar integrated photonic technology presents advantages such as high sensitivity, compactness, high multiplexing level, shorter time to result, need of very low sample and reagent volumes, immunity to electromagnetic interferences and compatibility with CMOS (Complementary Metal-Oxide-Semiconductor) technology fabrication [32][33][34]. Oligonucleotides detection has already been demonstrated using photonic based sensors such as ring resonators [35][36][37][38], photonic crystals [38,39], Mach-Zehnder interferometers [40,41] or bimodal waveguides [42], exhibiting outstanding performance.…”

Photonic biosensor chip for early-stage cancer diagnosis

“…Many methods have been explored for the detection of miRNAs (Deng, et al, 2014;Degliangeli, et al, 2014;Roy, et al, 2011;Driskell, et al, 2008), but they are mostly too expensive and time consuming, requiring specialised laboratory equipments, and tedious incubation steps (Wen et al, 2012). Research has also been carried out with implementing silicon nanowire technology and some progresses has been made (Qavi, et al, 2011;Densmore, et al, 2009;Pandraud,et al, 2000;Ymeti, et al, 2005;Zhang, 2013). The development of "lab on a chip" photonic biosensors with high sensitivity of detection, capable of detecting disease-associated miRNAs, would be beneficial as an effective tool for the early-stage diagnosis of various disesases, especially cancers, as miRNAs are well preserved in various human specimens, including urine (Waber et al, 2010).…”