Electrochemical analysis of HIV-1 reverse transcriptase serum level: exploiting protein binding to a functionalized...

Area of Science:

• Electrochemistry
• Biosensor Technology
• Biomolecular Detection

Background:

• Human immunodeficiency virus type-1 reverse transcriptase (HIV-1 RT) is a critical target for viral load monitoring and drug efficacy assessment.
• Existing detection methods may lack sensitivity, speed, or require complex sample preparation.
• Development of rapid, sensitive, and specific diagnostic tools for HIV-1 RT is essential for effective patient management.

Purpose of the Study:

• To develop and validate a novel electrochemical biosensor for the sensitive detection of HIV-1 RT in serum.
• To utilize an organometallic peptide conjugate immobilized on a nanostructured gold surface for enhanced signal transduction.
• To establish a quantitative assay with a low limit of detection and rapid response time.

Main Methods:

• Fabrication of a gold nanoparticles-functionalized screen-printed carbon electrode (GNPs-SPCE).
• Immobilization of ferrocene-labeled lipoic acid (Fc-LA) onto the electrode surface via a gold-thiol bond.
• Covalent attachment of a specific peptide (VEAIIRILQQLLFIH) to Fc-LA for HIV-1 RT recognition.
• Electrochemical detection using square wave voltammetry to measure changes in Fc redox signal upon RT binding.

Main Results:

• Successful formation of a stable Fc-LA thin film on the GNPs-SPCE, confirmed by TOF-SIMS and XPS.
• Development of a sensitive biosensor capable of detecting HIV-1 RT through electrochemical signal modulation.
• Linear quantification of HIV-1 RT in serum over a range of 1-500 pg mL⁻¹ (0.9-427 fM).
• Achieved a low limit of detection of 0.8 pg mL⁻¹ (0.7 fM) with a short response time.

Conclusions:

• The developed electrochemical biosensor provides a sensitive and rapid method for HIV-1 RT detection in serum.
• The organometallic peptide conjugate approach offers a promising platform for developing next-generation diagnostic tools for HIV-1.
• This assay demonstrates potential for clinical application in HIV monitoring and diagnostics.