Abstract
Nucleic acids and proteins are the two most important target types used in molecular diagnostics. In many instances, simultaneous sensitive and accurate detection of both biomarkers from the same sample would be desirable, but standard detection methods are highly optimized for one type and not cross-compatible. Here, we report the simultaneous multiplexed detection of SARS-CoV-2 RNAs and antigens with single molecule sensitivity. Both analytes are isolated and labeled using a single bead-based solid-phase extraction protocol, followed by fluorescence detection on a multi-channel optofluidic waveguide chip. Direct amplification-free detection of both biomarkers from nasopharyngeal swab samples is demonstrated with single molecule detection sensitivity, opening the door for ultrasensitive dual-target analysis in infectious disease diagnosis, oncology, and other applications.
Highlights
There is a pressing demand for tools that simultaneously analyze multiple biomarker types, such as nucleic acids, proteins, and metabolites
Multi-target analysis would raise reliability and confidence. Another example is the acute respiratory illness Coronavirus disease 2019 (COVID19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a highly infectious and harmful pathogen that attacks and destroys lung tissues
Both RT-PCR for viral RNA10 and enzyme linked immunosorbent assay (ELISA) and chemiluminescence immunoassays (CLIAs) for nucleocapsid antigens11 are used for COVID-19 confirmation
Summary
There is a pressing demand for tools that simultaneously analyze multiple biomarker types, such as nucleic acids, proteins, and metabolites This requirement is partially driven by the emergence of personalized medicine, single cell analysis, and the need to analyze a variety of genomic and proteomic biomarkers with high specificity and sensitivity—ideally at ultra-low concentrations for early disease detection.. We report the use of the ARROW photonic biosensor platform for the first dual detection of nucleic acid and antigen biomarkers with single molecule sensitivity from clinical SARSCoV-2 nasopharyngeal (NP) swab samples This is enabled by combining a bead-based extraction protocol, spatial–spectral multiplexing in a multi-channel chip, and novel bright fluorescent probes for protein targets in a single assay. We demonstrate amplificationfree detection of both targets with high accuracy using efficient wavelet-based data analysis
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