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Optofluidic multiplex detection of single SARS-CoV-2 and influenza A antigens using a novel bright fluorescent probe assay.
Stambaugh, Alexandra; Parks, Joshua W; Stott, Matthew A; Meena, Gopikrishnan G; Hawkins, Aaron R; Schmidt, Holger.
  • Stambaugh A; School of Engineering, University of California, Santa Cruz, CA 95064; amstamba@ucsc.edu.
  • Parks JW; School of Engineering, University of California, Santa Cruz, CA 95064.
  • Stott MA; Electrical and Computer Engineering Department, Brigham Young University, Provo, UT 84602.
  • Meena GG; School of Engineering, University of California, Santa Cruz, CA 95064.
  • Hawkins AR; Electrical and Computer Engineering Department, Brigham Young University, Provo, UT 84602.
  • Schmidt H; School of Engineering, University of California, Santa Cruz, CA 95064.
Proc Natl Acad Sci U S A ; 118(20)2021 05 18.
Article in English | MEDLINE | ID: covidwho-1216492
ABSTRACT
The urgency for the development of a sensitive, specific, and rapid point-of-care diagnostic test has deepened during the ongoing COVID-19 pandemic. Here, we introduce an ultrasensitive chip-based antigen test with single protein biomarker sensitivity for the differentiated detection of both severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza A antigens in nasopharyngeal swab samples at diagnostically relevant concentrations. The single-antigen assay is enabled by synthesizing a brightly fluorescent reporter probe, which is incorporated into a bead-based solid-phase extraction assay centered on an antibody sandwich protocol for the capture of target antigens. After optimization of the probe release for detection using ultraviolet light, the full assay is validated with both SARS-CoV-2 and influenza A antigens from clinical nasopharyngeal swab samples (PCR-negative spiked with target antigens). Spectrally multiplexed detection of both targets is implemented by multispot excitation on a multimode interference waveguide platform, and detection at 30 ng/mL with single-antigen sensitivity is reported.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Influenza A virus / Molecular Diagnostic Techniques / Microfluidic Analytical Techniques / SARS-CoV-2 / Antigens, Viral Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Year: 2021 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Influenza A virus / Molecular Diagnostic Techniques / Microfluidic Analytical Techniques / SARS-CoV-2 / Antigens, Viral Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Year: 2021 Document Type: Article