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A rapid, ultrasensitive voltammetric biosensor for determining SARS-CoV-2 spike protein in real samples.

ABSTRACT: The ongoing coronavirus disease 2019 (COVID-19) pandemic continues to threaten public health systems all around the world. In controlling the viral outbreak, early diagnosis of COVID-19 is pivotal. This article describes a novel method of voltammetrically determining severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein with a newly designed sensor involving bovine serum albumin, SARS-CoV-2 spike antibody and a functionalised graphene oxide modified glassy carbon electrode (BSA/AB/f-GO/GCE) or screen-printed electrode (BSA/AB/f-GO/SPE). The oxidation reaction based on the antibody-antigen protein interaction was evaluated as a response to SARS-CoV-2 spike protein at -200 mV and 1430 mV with the BSA/AB/f-GO/SPE and BSA/AB/f-GO/GCE, respectively. The developed sensors, BSA/AB/f-GO/SPE and BSA/AB/f-GO/GCE, could detect 1 ag/mL of virus spike protein in synthetic, saliva and oropharyngeal swab samples in 5 min and 35 min, and both sensors demonstrated a dynamic response to the SARS-CoV-2 spike protein between 1 ag/mL and 10 fg/mL. Real-time polymerase chain reaction (RT-PCR), rapid antigen test and the proposed method were applied to saliva samples. When compared to RT-PCR, it was observed that the developed method had a 92.5% specificity and 93.3% sensitivity. Moreover, BSA/AB/f-GO/SPE sensor achieved 91.7% accuracy compared to 66.7% accuracy of rapid antigen test kit in positive samples. In view of these findings, the developed sensor provides great potential for the diagnosing of COVID-19 in real samples.

SUBMITTER: Liv L

PROVIDER: S-EPMC8276568 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:A large-scale diagnosis of the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) is essential to downregulate its spread within as well as across communities and mitigate the current outbreak of the pandemic novel coronavirus disease 2019 (COVID-19). Herein, we report the development of a rapid (less than 5 min), low-cost, easy-to-implement, and quantitative paper-based electrochemical sensor chip to enable the digital detection of SARS-CoV-2 genetic material. The biosensor uses gold nanoparticles (AuNPs), capped with highly specific antisense oligonucleotides (ssDNA) targeting viral nucleocapsid phosphoprotein (N-gene). The sensing probes are immobilized on a paper-based electrochemical platform to yield a nucleic-acid-testing device with a readout that can be recorded with a simple hand-held reader. The biosensor chip has been tested using samples collected from Vero cells infected with SARS-CoV-2 virus and clinical samples. The sensor provides a significant improvement in output signal only in the presence of its target-SARS-CoV-2 RNA-within less than 5 min of incubation time, with a sensitivity of 231 (copies ?L-1)-1 and limit of detection of 6.9 copies/?L without the need for any further amplification. The sensor chip performance has been tested using clinical samples from 22 COVID-19 positive patients and 26 healthy asymptomatic subjects confirmed using the FDA-approved RT-PCR COVID-19 diagnostic kit. The sensor successfully distinguishes the positive COVID-19 samples from the negative ones with almost 100% accuracy, sensitivity, and specificity and exhibits an insignificant change in output signal for the samples lacking a SARS-CoV-2 viral target segment (e.g., SARS-CoV, MERS-CoV, or negative COVID-19 samples collected from healthy subjects). The feasibility of the sensor even during the genomic mutation of the virus is also ensured from the design of the ssDNA-conjugated AuNPs that simultaneously target two separate regions of the same SARS-CoV-2 N-gene.