Fluorescent bioassay for SARS-CoV-2 detection using polypyrene-g-poly(epsilon-caprolactone) prepared by simultaneous photoinduced step-growth and ring-opening polymerizations
Makaleler > Tam Makale · 2022
Yazarlar: Tugba Celiker, Faezeh Ghorbanizamani, Hichem Moulahoum, Emine GÜLER ÇELİK, Kerem Tok, Figen Zihnioglu, Candan Cicek, Ruchan Sertoz, Bilgin Arda, Tuncay Goksel, Suna TİMUR
MICROCHIMICA ACTA, SCI Expanded İndekslerine Giren Dergi, Uluslararası, 2022, cilt 189
DOI: 10.1007/s00604-022-05244-2
Özet
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.