Droplet Microarray Based on Nanosensing Probe Patterns for Simultaneous Detection of Multiple HIV Retroviral Nucleic Acids

Abstract

Multiplexed detection of viral nucleic acids is important for rapid screening of viral infection. In this study, we present a molybdenum disulfide (MoS 2 ) nanosheet-modified dendrimer droplet microarray (DMA) for rapid and sensitive detection of retroviral nucleic acids of human immunodeficiency virus-1 (HIV-1) and human immunodeficiency virus-2 (HIV-2) simultaneously. The DMA platform was fabricated by omniphobic–omniphilic patterning on a surface-grafted dendrimer substrate. Functionalized MoS 2 nanosheets modified with fluorescent dye-labeled oligomer probes were prepatterned on positively charged amino-modified omniphilic spots to form a fluorescence resonance energy transfer (FRET) sensing microarray. With the formation of separated microdroplets of sample on the hydrophobic–hydrophilic micropattern, prepatterned oligomer probes specifically hybridized with the target HIV genes and detached from the MoS 2 nanosheet surface due to weakening of the adsorption force, leading to fluorescence signal recovery. As a proof of concept, we used this microarray with a small sample size (<150 nL) for simultaneous detection of HIV-1 and HIV-2 nucleic acids with a limit of detection (LOD) of 50 pM. The multiplex detection capability was further demonstrated for simultaneous detection of five viral genes (HIV-1, HIV-2, ORFlab, and N genes of SARS-COV-2 and M gene of Influenza A). This work demonstrated the potential of this novel MoS 2 -DMA FRET sensing platform for high-throughput multiplexed viral nucleic acid screening.

Recent Advances in Two-Dimensional Transition Metal Dichalcogenide Nanocomposites Biosensors for Virus Detection before and during COVID-19 Outbreak

“…This microarray viral biosensor provides a high-throughput screening platform for simultaneous monitoring of multiplexed viral nucleic acids screening, such as the SARS-CoV-2 viruses, from a small sample size. During the outbreak of the COVID-19 pandemic in 2020, our research team has reported MoS 2 nanosheet-modified dendrimer droplet microarray (DMA) for simultaneously rapid and sensitive detection of five viral genes, HIV-1, HIV-2, ORFlab, and N genes of SARS-CoV-2 and M gene of Influenza A (Figure 9) [114]. MoS 2 nanosheets were conjugated onto cysteamine-containing DMA (MoS 2 -DMA), serving as acceptors substrate, and subsequently interacted with multiple fluorescent dye-labeled oligonucleotide probes serving as donors for FRET assays.…”

2D MOF Nanosensor‐Integrated Digital Droplet Microfluidic Flow Cytometry for In Situ Detection of Multiple miRNAs in Single CTC Cells

“…[17][18][19] The integration of microfluidics with nanosenosrs has led to the creation of low-cost lab-on-a-chip platforms for high-throughput bioanalysis. [20][21][22] Therefore, the integration of MOF-based nanosensors with droplet microfluidic devices can be a promising approach for in situ analysis of multiple miRNAs in single CTCs.…”

Programmable molecular circuit discriminates multidrug-resistant bacteria

“…Previous research has revealed that MoS 2 nanosheets could strongly adsorb single-stranded DNA (ssDNA) by van der Waals forces between nucleobases and the basal plane of MoS 2 nanosheets [ 39 , 40 ]. The fluorescence quenching capabilities of MoS 2 enable those fluorescent ssDNA probes loaded onto the MoS 2 nanosheets to construct a convenient biosensing strategy, which could correspond to the complementary ssDNA using an “off-to-on” signal readout mechanism [ [41] , [42] , [43] , [44] ]. Inspired by these previous reports, we proposed a b ackground-f r ee i sothermal c ircuital k it (BRICK) for the reliable discrimination of MDR bacteria.…”