A Cell-Based Electrochemical Biosensor for the Detection of Infectious Hepatitis A Virus

Area of Science:

• Virology
• Biosensor Technology
• Electrochemical Methods

Background:

• Hepatitis A virus (HAV) causes acute liver infections via the fecal-oral route.
• Current diagnostic methods detect HAV antigen or RNA but cannot distinguish infectious from non-infectious virus.
• There is a need for rapid and specific detection of infectious HAV.

Purpose of the Study:

• To develop a prototype cell-based electrochemical biosensor for detecting infectious HAV.
• To assess the biosensor's sensitivity, specificity, and stability.

Main Methods:

• Utilized a cell culture-adapted HAV strain (HM175/18f) and permissive cells (FRhK-4).
• Developed a biosensor using gold nanoparticle-modified screen-printed electrodes.
• Employed electrochemical impedance spectroscopy for signal quantification.

Main Results:

• Successfully fabricated the cell-based biosensor, confirmed by Nyquist plots.
• Achieved an optimal HAV incubation period of 6 hours.
• Demonstrated a linear relationship (R² = 0.98) with a limit of detection of approximately 5 TCID50/mL for infectious HAV.
• Showed no cross-reactivity with non-target viruses (feline calicivirus, human coronavirus 229E).
• Maintained 60-90% signal stability for 3-7 days at 37 °C.

Conclusions:

• The developed prototype cell-based biosensor can rapidly detect low levels of infectious HAV.
• This technology offers a potential advancement over existing methods by specifically identifying infectious viral particles.
• The biosensor shows promise for improved diagnostics and public health surveillance of Hepatitis A.