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An internal amplification control for quantitative nucleic acid analysis using nanoparticle-based dipstick

Huan Huang1, Li Jin, Xian Yang

  • 1Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China.

Biosensors & Bioelectronics
|December 5, 2012
PubMed
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This study introduces a novel nanoparticle-based DNA biosensor with an internal amplification control (IAC) for accurate virus nucleic acid quantification. This method enables sensitive detection of viral DNA, aiding in monitoring treatment efficacy.

Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Nanotechnology

Background:

  • Quantitative analysis of virus nucleic acids (RNA or DNA) in blood is crucial for assessing medical treatment effectiveness.
  • Current methods may require complex laboratory setups, limiting point-of-care applications.

Purpose of the Study:

  • To develop a quantitative method for detecting virus nucleic acids in blood using an internal amplification control (IAC) and a nanoparticle-based DNA biosensor.
  • To enable semi-quantitative analysis of viral load for improved point-of-care testing (POCT).

Main Methods:

  • An internal amplification control (IAC) with a specific sequence was designed and added to serum samples before nucleic acid extraction.
  • A dipstick biosensor utilizing streptavidin-functionalized nanoparticles was developed for visual detection of amplified targets.

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  • Amplicon detection was achieved by comparing color densities on the biosensor test and IAC zones.

    • Main Results:

    • The proposed method achieved detection of Hepatitis B virus (HBV) DNA at approximately 100 copies.
    • Successful semi-quantification of viral pathogen load was demonstrated in 30 clinical samples from HBV-infected patients.
    • The IAC and nanoparticle biosensor system showed high amplification efficiency, comparable to the target sequences.

    Conclusions:

    • The developed nanoparticle-based DNA biosensor coupled with an IAC offers a sensitive and feasible approach for semi-quantitative analysis of virus nucleic acids.
    • This method holds significant potential as a powerful tool for point-of-care testing (POCT) in clinical settings.
    • The system facilitates monitoring of viral load and treatment efficacy, particularly for infections like HBV.