Abstract
Detection and quantification of viral particles (VPs) facilitate both diagnostics of pathogenic viruses and quality control testing of virus-based products. However, existing technologies fail to afford concurrent ultrasensitive detection and large-scale absolute quantification of VPs. Here, we propose a digital Microgel-in-Droplet enzyme-linked immunosorbent assay (ELISA) system that enables the processing and monitoring of millions of ELISA reactions at the single-VP level by incorporating droplet microfluidics with sandwich ELISA. Upon validating the microfluidic workflow and optimizing ELISA parameters, we demonstrate ultrasensitive VP detection at a limit of detection of 56 PFU/test. Leveraging a fluorescence-based screening platform, we further realize high-throughput digital counting of VPs with a linear detection range of 500-64 000 PFU/test. The precision is comparable to that of the gold standard, the plaque assay, across a wide range of virus concentrations. We anticipate that our system will provide a novel paradigm for the absolute enumeration of various types of viral particles.
© 2024 American Chemical Society
© 2024 American Chemical Society
| Original language | English |
|---|---|
| Pages (from-to) | 16134-16144 |
| Journal | Analytical Chemistry |
| Volume | 96 |
| Issue number | 41 |
| Online published | 3 Oct 2024 |
| DOIs | |
| Publication status | Published - 15 Oct 2024 |
| Externally published | Yes |
Funding
The authors thank Prof. Leo Poon for kindly providing the virus samples. The authors acknowledge the financial support provided by the General Research Fund (17303123), Collaborative Research Fund (C7165-20GF) by the Research Grants Council of Hong Kong. L.N. and H.C.S. are also supported by the Health@InnoHK Program of the Innovation and Technology Commission of the Hong Kong SAR Government.
RGC Funding Information
- RGC-funded
