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Life sciences · Journal article
Sensors and Actuators Reports · November 1, 2026
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Rapid, selective nucleic acid detection is essential for environmental surveillance and public health monitoring. This study presents a label-free fluorescent biosensor based on polyaniline (PANI) functionalized with a DNA probe for SARS-CoV-2 RNA recognition. UV irradiation conditions and probe concentration were optimized to improve fluorescence performance. The biosensor discriminated complementary from non-complementary RNA sequences at concentrations as low as 10⁻¹⁰ mM. Concentration-dependent fluorescence quenching was observed over 10⁻¹⁰ – 10⁻⁶ mM, yielding a strong logarithmic calibration fit (R² = 0.9934). Blank-based limits of detection and quantification were statistically estimated at 1.83 × 10⁻⁶ and 6.11 × 10⁻⁶ mM, respectively, pending experimental validation. Principal component analysis and hierarchical clustering confirmed spectral discrimination between sensing stages and RNA concentrations. Electrochemical impedance spectroscopy, cyclic voltammetry, and ATR-FTIR provided complementary evidence of interfacial modifications following DNA functionalization and RNA hybridization. Sensor stability was additionally assessed over 21 days to investigate fluorescence-response reproducibility. Application to purified RNA extracts from municipal wastewater and spent hemodialysis dialysate yielded relative fluorescence-response amplitudes of 92.3% and 15.1%, respectively, indicating distinct matrix-dependent effects rather than concentration-based analytical recoveries. Comparison with RT-qPCR measurements showed a strong association (R² = 0.9025), supporting the platform’s applicability to environmental samples. Overall, the PANI–DNA biosensor offers a promising amplification-free sensing strategy for SARS-CoV-2 RNA recognition and wastewater-based epidemiology, although further validation of analytical sensitivity, matrix tolerance, and operational robustness remains necessary.