A Phase Separation-Assisted Pre-Enrichment Method for Ultrasensitive Respiratory Virus Detection

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Biochemistry

Background:

• Biomedical applications require enrichment of trace biomarkers like proteins and nucleic acids.
• Conventional enrichment methods often lack versatility for diverse biomarker types.

Purpose of the Study:

• To introduce and optimize phase separation-assisted pre-enrichment (PSAP) technology for versatile biomarker enrichment.
• To evaluate the performance of PSAP in enhancing the detection of viral antigens and RNA.

Main Methods:

• Exploiting differential polymer-polymer partitioning for simultaneous antigen and RNA enrichment.
• Systematic optimization of interfacial chemistry (pH, polymer properties) for protocol refinement.
• Integration of PSAP with commercial diagnostics, including rapid antigen tests (RATs).

Main Results:

• Achieved 47-fold antigen and 44-fold RNA enrichment simultaneously using PSAP.
• PSAP-boosted RATs showed tenfold lower limits for SARS-CoV-2 and fivefold for Influenza.
• Significantly enhanced detection accuracy and positivity rates in low viral load specimens, maintaining specificity.

Conclusions:

• PSAP technology offers universal applicability and tunability for enriching diverse biomarkers.
• PSAP enhances sensitivity and accuracy in point-of-care diagnostics for respiratory pathogens.
• PSAP provides a foundation for next-generation diagnostic tools.