Fluorescent bioassay for SARS-CoV-2 detection using polypyrene-g-poly(epsilon-caprolactone) prepared by simultaneous photoinduced step-growth and ring-opening polymerizations


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Celiker T., Ghorbanizamani F., Moulahoum H., GÜLER ÇELİK E., TOK K., ZİHNİOĞLU F., ...More

MICROCHIMICA ACTA, vol.189, no.5, 2022 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 189 Issue: 5
  • Publication Date: 2022
  • Doi Number: 10.1007/s00604-022-05244-2
  • Title of Journal : MICROCHIMICA ACTA
  • Keywords: Photopolymerization, Step-growth polymerization, Ring-opening polymerization, Graft copolymer, Fluorescent biosensor, SARS-CoV-2, LATERAL FLOW ASSAY, POLYMERSOMES, DIAGNOSIS

Abstract

The construction of a rapid and easy immunofluorescence bioassay for SARS-CoV-2 detection is described. We report for the first time a novel one-pot synthetic approach for simultaneous photoinduced step-growth polymerization of pyrene (Py) and ring-opening polymerization of epsilon-caprolactone (PCL) to produce a graft fluorescent copolymer PPy-g-PCL that was conjugated to SARS-CoV-2-specific antibodies using EDC/NHS chemistry. The synthesis steps and conjugation products were fully characterized using standard spectral analysis. Next, the PPy-g-PCL was used for the construction of a dot-blot assay which was calibrated for applications to human nasopharyngeal samples. The analytical features of the proposed sensor showed a detection range of 6.03-8.7 LOG viral copy mL(-1) (Ct Scores: 8-25), the limit of detection (LOD), and quantification (LOQ) of 1.84 and 6.16 LOG viral copy mL(-1), respectively. The repeatability and reproducibility of the platform had a coefficient of variation (CV) ranging between 1.2 and 5.9%. The fluorescence-based dot-blot assay was tested with human samples. Significant differences were observed between the fluorescence intensity of the negative and positive samples, with an overall correct response of 93.33%. The assay demonstrated a high correlation with RT-PCR data. This strategy opens new insights into simplified synthesis procedures of the reporter molecules and their high potential sensing and diagnosis applications.