ABSTRACT
COVID-19 has spread worldwide and early detection has been the key to controlling its propagation and preventing severe cases. However, diagnostic devices must be developed using different strategies to avoid a shortage of supplies needed for tests' fabrication caused by their large demand in pandemic situations. Furthermore, some tropical and subtropical countries are also facing epidemics of Dengue and Zika, viruses with similar symptoms in early stages and cross-reactivity in serological tests. Herein, we reported a qualitative immunosensor based on capacitive detection of spike proteins of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of COVID-19. The sensor device exhibited a good signal-to-noise ratio (SNR) at 1 kHz frequency, with an absolute value of capacitance variation significantly smaller for Dengue and Zika NS1 proteins (|ΔC| = 1.5 ± 1.0 nF and 1.8 ± 1.0 nF, respectively) than for the spike protein (|ΔC| = 7.0 ± 1.8 nF). Under the optimized conditions, the established biosensor is able to indicate that the sample contains target proteins when |ΔC| > 3.8 nF, as determined by the cut-off value (CO). This immunosensor was developed using interdigitated electrodes which require a measurement system with a simple electrical circuit that can be miniaturized to enable point-of-care detection, offering an alternative for COVID-19 diagnosis, especially in areas where there is also a co-incidence of Zika and Dengue.
ABSTRACT
The early diagnosis of Coronavirus disease (COVID-19) requires either an accurate detection of genetic material or a sensitive detection of viral proteins. In this work, we designed an immunoassay platform for detecting trace levels of SARS-CoV-2 spike (S) protein. It is based on surface-enhanced resonance Raman scattering (SERRS) of methylene blue (MB) adsorbed onto spherical gold nanoparticles (AuNPs) and coated with a 6 nm silica shell. The latter shell in the SERRS nanoprobe prevented aggregation and permitted functionalization with SARS-CoV-2 antibodies. Specificity of the immunoassay was achieved by combining this functionalization with antibody immobilization on the cover slides that served as the platform support. Different concentrations of SARS-CoV-2 antigen could be distinguished and the lack of influence of interferents was confirmed by treating SERRS data with the multidimensional projection technique Sammon's mapping. With SERRS using a laser line at 633 nm, the lowest concentration of spike protein detected was 10 pg/mL, achieving a limit of detection (LOD) of 0.046 ng/mL (0.60 pM). This value is comparable to the lowest concentrations in the plasma of COVID-19 patients at the onset of symptoms, thus indicating that the SERRS immunoassay platform may be employed for early diagnosis.