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Thermally Triggered Double Emulsion-Integrated Hydrogel Microparticles for Multiplexed Molecular Diagnostics
Eui Ju Jeon1,2, Seungwon Jung1,3, Yoon-Ha Jang1
1Center for Advanced Biomolecular Recognition, Biomedical Research Division, Korea Institute of Science and Technology (KIST), Seoul, 02792, Republic of Korea.
Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|January 17, 2025
Summary
A novel multiplexed assay, TaqPIN RT-qPCR, enables simultaneous detection of multiple genes, including SARS-CoV-2 variants, using a single optical channel for improved diagnostics.
Area of Science:
- Molecular Biology
- Biotechnology
- Diagnostic Assays
Background:
- Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is a key diagnostic tool for COVID-19.
- Current RT-qPCR assays are typically single-plex, limiting simultaneous gene analysis.
- Multiplexed assays are needed to expand the applications of RT-qPCR.
Purpose of the Study:
- To develop a novel multiplexed RT-qPCR assay for simultaneous detection of multiple genes.
- To enhance the capabilities of RT-qPCR for pathogen detection and variant discrimination.
- To create a single-channel multiplexed assay using microparticle technology.
Main Methods:
- Development of primer-incorporated network tailored with Taqman probe (TaqPIN) RT-qPCR.
- Utilizing a double emulsion (DE)-based carrier and polymer microparticle reactor.
- Loading multiple microparticles into a single reaction for multiplexing.
Main Results:
- TaqPIN RT-qPCR achieved 93.8% amplification efficiency and detected down to 20 copies/µL.
- A nine-plex assay successfully distinguished between SARS-CoV-2 variants, detecting single nucleotide variations.
- Testing on 75 samples showed 100% sensitivity and specificity for SARS-CoV-2 detection and 94% accuracy for variant discrimination.
Conclusions:
- The developed TaqPIN RT-qPCR is a sensitive and specific multiplexed assay.
- This technology enables single-channel multiplexing for advanced genetic analysis.
- The assay shows significant potential for accurate pathogen detection and variant tracking.
