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<p>There is a pressing need for a new generation of diagnostic technologies compatible with the combined characteristics of low-cost instrumentation, simple assay methods, high sensitivity, accurate quantitation, and rigorous validity. In this work, we introduce a plasmonic fluor-linked immunosorbent assay (p-FLISA) that enables sensitive and accurate detection and quantification of viral RNA in samples obtained from COVID-19 patients. The assay relies on a monoclonal antibody (S9.6) that specifically binds to DNA-RNA heteroduplexes comprising viral RNA and complementary DNA. The p-FLISA exhibited more than 200-fold lower limit-of-detection (LOD) and 70-fold lower limit-of-quantification (LLOQ) than the gold standard enzyme-linked immunosorbent assay (ELISA). Implementing the immunoassay in digital format (i.e., counting the individual fluorescent nanolabels instead of measuring the ensemble fluorescence intensity) resulted in a nearly 2300-fold improvement in the LOD and 460-fold improvement in the LLOQ compared to ELISA. The clinical sensitivity and specificity of this simple, scalable, and amplification-free assay are comparable to reverse transcription-polymerase chain reaction (RT-PCR). However, as opposed to RT-PCR, the assay enables the absolute quantification of RNA concentration, potentially enabling stratification of the stage of illness and infectivity of the patients. The novel method demonstrated here can be easily adapted for amplification-free detection and quantification of various RNA targets.</p>

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Plasmon-enhanced bioassay for amplification-free detection and quantification of SARS-CoV-2 RNA

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年2月19日

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  • Lin Liu,
  • Anushree Seth,
  • Yuxiong Liu,
  • Rohit Gupta,
  • Zheyu Wang,
  • Yixuan Wang,
  • Priya Rathi,
  • Gayathri Kondepati,
  • Jeremiah J. Morrissey,
  • Gregory Storch,
  • Guy M. Genin,
  • Bijal Parikh,
  • Sumanth Gandra,
  • Ige George,
  • Srikanth Singamaneni

摘要

There is a pressing need for a new generation of diagnostic technologies compatible with the combined characteristics of low-cost instrumentation, simple assay methods, high sensitivity, accurate quantitation, and rigorous validity. In this work, we introduce a plasmonic fluor-linked immunosorbent assay (p-FLISA) that enables sensitive and accurate detection and quantification of viral RNA in samples obtained from COVID-19 patients. The assay relies on a monoclonal antibody (S9.6) that specifically binds to DNA-RNA heteroduplexes comprising viral RNA and complementary DNA. The p-FLISA exhibited more than 200-fold lower limit-of-detection (LOD) and 70-fold lower limit-of-quantification (LLOQ) than the gold standard enzyme-linked immunosorbent assay (ELISA). Implementing the immunoassay in digital format (i.e., counting the individual fluorescent nanolabels instead of measuring the ensemble fluorescence intensity) resulted in a nearly 2300-fold improvement in the LOD and 460-fold improvement in the LLOQ compared to ELISA. The clinical sensitivity and specificity of this simple, scalable, and amplification-free assay are comparable to reverse transcription-polymerase chain reaction (RT-PCR). However, as opposed to RT-PCR, the assay enables the absolute quantification of RNA concentration, potentially enabling stratification of the stage of illness and infectivity of the patients. The novel method demonstrated here can be easily adapted for amplification-free detection and quantification of various RNA targets.