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

Yang Cao; Pui Ngan Lau; Alex W. H. Chin; Zhuolin He; Christina C. K. Au Yeung; Kehao Zeng; Haisong Lin; Leo L. M. Poon; Ho Cheung Shum

doi:10.1002/advs.202506578

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

Yang Cao, Pui Ngan Lau, Alex W. H. Chin, Zhuolin He, Christina C. K. Au Yeung, Kehao Zeng, Haisong Lin^*, Leo L. M. Poon^*, Ho Cheung Shum^*

^*Corresponding author for this work

Research output: Journal Publications and Reviews › RGC 21 - Publication in refereed journal › peer-review

2 Citations (Scopus)

33 Downloads (CityUHK Scholars)

Abstract

Enriching trace biomarkers (e.g., proteins, nucleic acids) is critical for biomedical applications; yet conventional methods often lack versatility, limiting their effectiveness to specific biomarker types. To address this, the phase separation-assisted pre-enrichment (PSAP) technology is presented that exploits differential polymer-polymer partitioning to achieve 47-fold antigen and 44-fold RNA enrichment simultaneously. Through systematic optimization of interfacial chemistry, including pH modulation, polymer hydrophilicity, mass fraction, and molecular weights, the protocol is refined to enable direct integration with commercial diagnostics. PSAP-boosted rapid antigen ests (RATs) detected SARS-CoV-2 and Influenza viruses at tenfold and fivefold lower limits, respectively. In clinical validation, 53 clinical specimens (containing PCR undetectable samples as controls) are analyzed. The PSAP method significantly enhanced detection accuracy for both viral antigens and RNA, particularly improving positivity rates in low viral load specimens (27 < Ct < 31) compared to conventional approaches while maintaining specificity in high-Ct and negative controls. With its universality and tunability, PSAP demonstrates universal applicability across respiratory pathogens and lays the foundation for next-generation point-of-care diagnostics. © 2025 The Author(s). Advanced Science published by Wiley-VCH GmbH

Original language	English
Article number	e06578
Journal	Advanced Science
DOIs	https://doi.org/10.1002/advs.202506578
Publication status	Online published - 27 Jun 2025

Funding

This study was supported by the Research Grants Council (RGC) of Hong Kong through the Collaborative Research Fund (C7165-20GF). This study was funded in part by the Health@InnoHK initiative of the Innovation and Technology Commission of the Hong Kong Special Administrative Region Government. It was partially supported by the Research Grants Council of HK theme-based research schemes (T11-705/21-N). The study was supported by the Hong Kong Jockey Club Global Health Institute (HKJCGHI), Hong Kong Special Administrative Region, China. H.C.S. was funded in part by the RGC Senior Research Fellow (SRFS2425-7S04) by the RGC. Haisong Lin acknowledges support from the NSFC Young Scientists Fund (Hong Kong) (No. 32201181) and the General Research Fund (No. 17208623). The authors thank Mr. Alvin Wong from Stanford University, who helped to measure the viscosity of dextran solutions during his summer internship. The authors thank Dr. Jingxuan Tian, Dr. Ruotong Zhang, Dr. Huanqing Cui, Dr. Qingchuan Li, and Mr. Chengzhi Zhang for their helpful discussions and suggestions.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Research Keywords

affinity-driven partitioning
aqueous two-phase systems
phase separation-assisted preconcentration
point-of-care testing
respiratory virus detection

Publisher's Copyright Statement

This full text is made available under CC-BY 4.0. https://creativecommons.org/licenses/by/4.0/

RGC Funding Information

RGC-funded

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abstract = "Enriching trace biomarkers (e.g., proteins, nucleic acids) is critical for biomedical applications; yet conventional methods often lack versatility, limiting their effectiveness to specific biomarker types. To address this, the phase separation-assisted pre-enrichment (PSAP) technology is presented that exploits differential polymer-polymer partitioning to achieve 47-fold antigen and 44-fold RNA enrichment simultaneously. Through systematic optimization of interfacial chemistry, including pH modulation, polymer hydrophilicity, mass fraction, and molecular weights, the protocol is refined to enable direct integration with commercial diagnostics. PSAP-boosted rapid antigen ests (RATs) detected SARS-CoV-2 and Influenza viruses at tenfold and fivefold lower limits, respectively. In clinical validation, 53 clinical specimens (containing PCR undetectable samples as controls) are analyzed. The PSAP method significantly enhanced detection accuracy for both viral antigens and RNA, particularly improving positivity rates in low viral load specimens (27 < Ct < 31) compared to conventional approaches while maintaining specificity in high-Ct and negative controls. With its universality and tunability, PSAP demonstrates universal applicability across respiratory pathogens and lays the foundation for next-generation point-of-care diagnostics. {\textcopyright} 2025 The Author(s). Advanced Science published by Wiley-VCH GmbH",

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AU - Cao, Yang

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AU - Chin, Alex W. H.

AU - He, Zhuolin

AU - Yeung, Christina C. K. Au

AU - Zeng, Kehao

AU - Lin, Haisong

AU - Poon, Leo L. M.

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ER -

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

Abstract

Funding

UN SDGs

Research Keywords

Publisher's Copyright Statement

RGC Funding Information

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Permanent Link

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SRFS: Deciphering Sequence-Dependent Molecular Interactions in Oligonucleotide-Based Coacervates: A Paradigm Shift in Biomaterial Design and Biomedical Engineering

Cite this

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

Abstract

Funding

UN SDGs

Research Keywords

Publisher's Copyright Statement

RGC Funding Information

Access Output

Other Access Options

Permanent Link

Other Files and Links

Fingerprint

Projects

SRFS: Deciphering Sequence-Dependent Molecular Interactions in Oligonucleotide-Based Coacervates: A Paradigm Shift in Biomaterial Design and Biomedical Engineering

Cite this