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Fluorescent bioassay for SARS-CoV-2 detection using polypyrene-g-poly(ε-caprolactone) prepared by simultaneous photoinduced step-growth and ring-opening polymerizations.
Celiker, Tugba; Ghorbanizamani, Faezeh; Moulahoum, Hichem; Guler Celik, Emine; Tok, Kerem; Zihnioglu, Figen; Cicek, Candan; Sertoz, Ruchan; Arda, Bilgin; Goksel, Tuncay; Turhan, Kutsal; Timur, Suna; Yagci, Yusuf.
  • Celiker T; Department of Chemistry, Istanbul Technical University, 34469, Maslak, Istanbul, Turkey.
  • Ghorbanizamani F; Department of Biochemistry, Faculty of Science, Ege University, 35100, Bornova, Izmir, Turkey.
  • Moulahoum H; Department of Biochemistry, Faculty of Science, Ege University, 35100, Bornova, Izmir, Turkey.
  • Guler Celik E; Department of Bioengineering, Faculty of Engineering, Ege University, 35100, Bornova, Izmir, Turkey. emine.guler.celik@ege.edu.tr.
  • Tok K; Department of Biochemistry, Faculty of Science, Ege University, 35100, Bornova, Izmir, Turkey.
  • Zihnioglu F; Department of Biochemistry, Faculty of Science, Ege University, 35100, Bornova, Izmir, Turkey.
  • Cicek C; Department of Medical Microbiology, Faculty of Medicine, Ege University, 35100, Bornova, Izmir, Turkey.
  • Sertoz R; Department of Medical Microbiology, Faculty of Medicine, Ege University, 35100, Bornova, Izmir, Turkey.
  • Arda B; Department of Infectious Diseases and Clinical Microbiology, Faculty of Medicine, Ege University, 35100, Bornova, Izmir, Turkey.
  • Goksel T; Department of Pulmonary Medicine, Faculty of Medicine, Ege University, 35100, Bornova, Izmir, Turkey.
  • Turhan K; EGESAM-Ege University Translational Pulmonary Research Center, 35100, Bornova, Izmir, Turkey.
  • Timur S; Department of Thoracic Surgery, Faculty of Medicine, Ege University, 35100, Bornova, Izmir, Turkey.
  • Yagci Y; Department of Biochemistry, Faculty of Science, Ege University, 35100, Bornova, Izmir, Turkey.
Mikrochim Acta ; 189(5): 202, 2022 04 26.
Article in English | MEDLINE | ID: covidwho-1850343
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 ε-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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Mikrochim Acta Year: 2022 Document Type: Article Affiliation country: S00604-022-05244-2

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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Mikrochim Acta Year: 2022 Document Type: Article Affiliation country: S00604-022-05244-2