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A General Biosensing Strategy Based on Cascade Amplification for Enhanced HIV Detection Sensitivity.

Yirui Zhang1,2, Jieyu Yan2, Wangheng Hou3

  • 1School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, China.

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A novel biosensing strategy uses a triple amplification cascade for highly sensitive detection of disease biomarkers like HIV-1 DNA and p24. This versatile platform shows promise for rapid molecular diagnostics in healthcare.

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Area of Science:

  • Biomolecular detection
  • Molecular diagnostics
  • Biosensor technology

Background:

  • Accurate and rapid detection of disease biomarkers is crucial for early diagnosis and effective management.
  • Current diagnostic methods may lack the sensitivity or speed required for timely intervention.
  • Acquired immunodeficiency syndrome (AIDS) biomarkers, such as HIV-1 DNA and p24 protein, require sensitive detection.

Purpose of the Study:

  • To propose a general biosensing strategy utilizing a treble signal amplification cascade.
  • To demonstrate the versatility and applicability of this strategy for diverse biomolecular targets.
  • To develop sensitive biosensors for detecting HIV-1 DNA and HIV-1 p24.

Main Methods:

  • A cascade amplification approach integrating isothermal amplification, rolling circle amplification (RCA), and CRISPR-Cas12a trans-cleavage.
  • Initial amplification triggered by probe hybridization and enzymatic activity.
  • Subsequent amplification steps via RCA and CRISPR-Cas12a for tertiary signal enhancement.

Main Results:

  • Achieved remarkable sensitivity with detection limits of 62 aM for nucleic acids and 8.48 pg/ml for proteins.
  • Demonstrated the platform's capability in clinical diagnosis through assays on clinical samples.
  • Validated the strategy's effectiveness for detecting HIV-1 DNA and HIV-1 p24.

Conclusions:

  • The cascade amplification strategy offers a broadly applicable framework for molecular diagnostics.
  • The modular design allows adaptation for various biomarkers beyond HIV.
  • This technology holds significant potential as a universal platform for healthcare and research.