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Dual-Gene-Controlled Rolling Circle Amplification Strategy for SARS-CoV-2 Analysis.

Ying Deng1, Tianci Zhou1, Ying Peng1

  • 1State Key Laboratory of Analytical Chemistry for Life Science, School of Life Sciences, Nanjing University, Nanjing 210023, P. R. China.

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|February 1, 2023
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Summary

This study presents a novel electrochemical method for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The dual-gene-controlled assay accurately identifies the virus by amplifying specific gene sequences, offering a sensitive diagnostic tool.

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Area of Science:

  • Biotechnology
  • Biosensors
  • Molecular Diagnostics

Background:

  • Simultaneous detection of multiple nucleic acids remains challenging.
  • Accurate and sensitive methods are crucial for diagnosing viral infections like COVID-19.

Purpose of the Study:

  • To develop a sensitive and accurate electrochemical method for simultaneous detection of two SARS-CoV-2 genes.
  • To utilize a dual-gene-controlled rolling circle amplification (RCA) strategy for enhanced detection.

Main Methods:

  • Designed a dual-gene-controlled RCA strategy targeting the N gene and RdRp gene of SARS-CoV-2.
  • Generated G-quadruplex (G4)-forming sequences triggering amplified electrochemical signals via G4/hemin complexes.
  • Employed TMB/H2O2 substrates for signal amplification and detection.

Main Results:

  • Achieved sensitive detection of SARS-CoV-2 with a limit of 57 fM in the range of 0.1-5000 pM.
  • Demonstrated high specificity by requiring the simultaneous presence of both target genes, preventing false positives.
  • Showed excellent agreement with RT-PCR results in distinguishing COVID-19 samples from healthy individuals.

Conclusions:

  • The label-free, dual-gene-controlled RCA strategy offers a highly sensitive and specific electrochemical method for SARS-CoV-2 detection.
  • This approach shows significant potential for accurate clinical diagnosis of COVID-19.
  • The method's accuracy and sensitivity make it a promising tool for infectious disease monitoring.