4.8 Article

Sensitive and specific clinically diagnosis of SARS-CoV-2 employing a novel biosensor based on boron nitride quantum dots/flower-like gold nanostructures signal amplification

Journal

BIOSENSORS & BIOELECTRONICS
Volume 207, Issue -, Pages -

Publisher

ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2022.114209

Keywords

COVID-19 pandemic; Virus diagnosis; SARS-CoV-2 RNA; Electrochemical biosensor

Funding

  1. Iran National Science Foundation (INSF) [99004067]
  2. Mashhad University of Medical Sciences, Mashhad, Iran [990524]

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This study presents a functionalized carbon electrode with specific antisense DNA oligonucleotide as a promising biosensor for targeting SARS-CoV-2 RNA without nucleic acid amplification. The biosensor showed excellent sensitivity and specificity in detecting the virus, with a lower detection limit compared to RT-PCR. It also exhibited negligible cross-reactivity with other viruses and remained stable for an extended period.
The sudden increase of the COVID-19 outbreak and its continued growth with mutations in various forms has created a global health crisis as well as devastating social and economic effects over the past two years. In this study, a screen-printed carbon electrode reinforced with boron nitride quantum dots/flower-like gold nano structures (BNQDs/FGNs/SPCE) and functionalized by highly specific antisense DNA oligonucleotide presents an alternative and promising solution for targeting SARS-CoV-2 RNA without nucleic acid amplification. The platform was tested on 120 SARS-CoV-2 RNA isolated from real clinical samples (60 positive and 60 negative confirmed by conventional RT-PCR method). Based on obtained quantitative results and statistical analysis (box diagram, cutoff value, receiver operating characteristic curve, and t-test), the biosensor revealed a significant difference between the two positive and negative groups with 100% sensitivity and 100% specificity. To evaluate the quantitation capacity and detection limit of the biosensor for clinical trials, the detection performance of the biosensor for continuously diluted RNA isolated from SARS-CoV-2-confirmed patients was compared to those obtained by RT-PCR, demonstrating that the detection limit of the biosensor is lower than or comparable to that of RT-PCR. The ssDNA/BNQDs/FGNs/SPCE showed negligible cross-reactivity with RNA fragments isolated from Influenza A (IAV) clinical samples and also remained stable for up to 14 days. In conclusion, the fabricated biosensor may serve as a promising tool for point-of-care applications.

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