Multiplex detection of blood-borne pathogens on a self-driven microfluidic chip using loop-mediated isothermal...

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Public Health

Background:

• Accurate detection of blood-borne pathogens, including hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV), is critical for blood transfusion safety.
• Conventional PCR methods for pathogen nucleic acid detection require specialized equipment, limiting their use in resource-constrained environments.

Purpose of the Study:

• To develop a self-driven microfluidic chip for multiplex loop-mediated isothermal amplification (LAMP) detection of HBV, HCV, and HIV.
• To assess the chip's performance, including sample loading time, detection sensitivity, and clinical applicability, particularly for resource-limited areas.

Main Methods:

• A self-driven microfluidic chip utilizing polydimethylsiloxane (PDMS) was designed for multiplex LAMP.
• Sample loading was achieved via pressure differences within 12 minutes, eliminating the need for external pumps.
• Multiplex detection was enabled by pre-setting specific LAMP primers in different reaction chambers, with calcein used as a visual indicator.

Main Results:

• The chip successfully detected as low as 2 copies/μL of HBV, HCV, and HIV target nucleic acids after 50 minutes of isothermal amplification at 63°C.
• Positive amplification was visually indicated by calcein, with results recordable via smartphone camera.
• Detection results for HBV in 20 clinical plasma samples showed consistency with a standard qPCR-based kit.

Conclusions:

• The developed self-driven microfluidic chip offers a promising, pump-free solution for multiplex blood-borne pathogen detection.
• This technology demonstrates significant potential for clinical application, especially in resource-limited settings, enhancing blood transfusion safety.
• The system's ease of use and smartphone compatibility facilitate accessible diagnostics.