DEVELOPMENT OF REDUCED GRAPHENE OXIDE (RGO) NANOSHEETS FOR ULTRASENSITIVE DETECTION OF A NOVEL CORONA VIRUS SARS-COV-2

DEEPAK V. SAWANT; PRANAV K. KATKAR; ARUNKUMAR PARTHASARATHY; R. K. SHARMA; SHIVAJI KASHTE; C. D. LOKHANDE

doi:10.22159/ijap.2025v17i5.53365

Authors

DEEPAK V. SAWANT Department of Microbiology, Dr. D. Y. Patil Medical College Hospital and Research Institute Kolhapur-416006, Maharashtra, India
PRANAV K. KATKAR Department of Physics, Sejong University, Gwangjin-gu, Seoul-05006, Republic of Korea
ARUNKUMAR PARTHASARATHY Department of Microbiology, Dr. D. Y. Patil Medical College Hospital and Research Institute Kolhapur-416006, Maharashtra, India
R. K. SHARMA Department of Microbiology, Dr. D. Y. Patil Medical College Hospital and Research Institute Kolhapur-416006, Maharashtra, India
SHIVAJI KASHTE Centre for Interdisciplinary Research, D. Y. Patil Education Society (Deemed to be University), Kolhapur-416006, Maharashtra, India
C. D. LOKHANDE Centre for Interdisciplinary Research, D. Y. Patil Education Society (Deemed to be University), Kolhapur-416006, Maharashtra, India

DOI:

https://doi.org/10.22159/ijap.2025v17i5.53365

Keywords:

Reduced graphene oxide, Ribonucleic acid, SARS-Cov-2, COVID-19 detection, Nano-sheets

Abstract

Objective: The extraction of ribonucleic acid (RNA) is essential for SARS-CoV-2 RT-PCR detection. Although commercial RNA extraction kits are available, they suffer from limitations such as low RNA yield, difficulty in amplification during RT-PCR, and high costs. This study investigates the evolution of rGO Nano sheets as a novel material for efficient RNA extraction from clinical specimens. The rGO Nano sheets offer a promising alternative for the purification of excellent RNA, which is crucial for accurate and high-sensitivity identification of SARS-CoV-2. By leveraging a unique property of rGO, the research demonstrates enhanced RNA recovery, improving the overall sensitivity and reliability of detection method for SARS-CoV-2.

Methods: The rGO Nano sheets were synthesized by reducing graphene oxide (GO) using a simple hydrothermal process. The rGO analysis and its interaction with RNA were characterized using X-ray diffraction, Raman spectroscopy, field emission scanning electron microscopy (FE-SEM), and Brunauer-Emmett-Teller (BET) analysis. RNA was extracted from clinical specimens using rGO Nano sheets via adsorption and elution and compared with extraction performed using a commercial RNA kit. RNA extraction efficiency and validation were assessed by UV-visible spectrometry and RT-PCR.

Results: The rGO Nano sheets exhibited a multi-layered carbon nanostructure with an average 7.15 nm 3D porous structure, a large surface area of 225 m²/g, and interconnected carbon Nano sheets. The RNA extracted using rGO Nano sheets was validated by UV-visible spectrometry and RT-PCR, showing successful detection of viral genes (ORF1ab and N gene) with a sensitivity of 10 copies. Ten to twenty copies of SARS-CoV-2 pseudo-virus particles showed a strong linear association. The P-value is greater than 0.05, indicating acceptance of the null hypothesis. The commercial kit and the rGO Nano sheet-based RNA extraction methods are not significantly different from one another. There is a significant increase in RNA yield (1.959 ng/µl) and improved sensitivity in RT-PCR, further validating the advantage of using rGO for RNA extraction.

Conclusion: The developed rGO Nano sheet-based RNA extraction method demonstrates high sensitivity and efficiency, offering a promising alternative to commercial kits for the detection of SARS-CoV-2.

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