Summary:
UCLA researchers in the Department of Bioengineering have developed a novel method to perform ultra-high sensitivity, low-cost nucleic acid amplification assays utilizing porous microdroplets.
Background:
Nucleic acid concentration measurement is essential in biological research and diagnostics. The 2020 pandemic underscored the importance of accessible and affordable tests to determine the presence of specific viruses or bacteria. These tests are used not only for infectious diseases but also in detecting cancers and monitoring public health, such as checking for pathogens in water sources. Traditionally, bulk assay methods like qPCR or next generation sequencing have been employed, but they have limitations in accuracy and cost. Recent advancements introduced digital nucleic acid tests (dNAATs), offering more precise results by analyzing individual molecules in tiny volumes. These digital assays can detect even low-abundance targets, vital for certain diagnostics like cancer. However, the current gold-standard technologies, like lab well plates, have constraints regarding the range of concentrations they can measure. New platforms have addressed these limitations; digital droplets and microwells increase dynamic range but suffer from throughput and cost issues while requiring specialized equipment.
Innovation:
To address current limitations in nucleic acid quantification, UCLA researchers have developed a microparticle-based platform for ultra-high throughput dNAATs. Unlike traditional methods, these hollow particles enable the creation of thousands of uniform partitions without the use of any additional specialized equipment. A hallmark of the technology is its unparalleled dynamic range, with ultra-low concentration detection capabilities, enabling particularly accurate nucleic acid measurements. Its ability to retain high-molecular weight DNA concatemers within the particles simplifies downstream analysis; These particles are easily analyzed using commercial flow cytometers commonly found in research labs, making the technology more accessible and cost-effective. Additionally, reagent use and minimum volumes are decreased, further driving the cost of nucleic acid assays down. With this platform, the inventors have paved the way for more precise and efficient nucleic acid diagnostics.
Potential Applications:
• Rapid Diagnostics
• Environmental monitoring & public health
• High-throughput genetic screening
• Biomanufacturing quality control
Advantages:
• Cost effective
• High sensitivity
• Extended dynamic range
• Broad accessibility
Status of Development:
The inventors have developed the microparticle system and successfully demonstrated the proof of concept in the laboratory.
Related Publications:
1. Wang, Y., Shah, V., Lu, A., Pachler, E., Cheng, B. and Di Carlo, D., 2021. Counting of enzymatically amplified affinity reactions in hydrogel particle-templated drops. Lab on a Chip, 21(18), pp.3438-3448.
2. van Zee, M., de Rutte, J., Rumyan, R., Williamson, C., Burnes, T., Radakovits, R., Sonico Eugenio, A., Badih, S., Lee, S., Lee, D.H. and Archang, M., 2022. High-throughput selection of cells based on accumulated growth and division using PicoShell particles. Proceedings of the National Academy of Sciences, 119(4), p.e2109430119.
Reference:
UCLA Case No. 2023-058
Lead Inventor:
Dino DiCarlo, UCLA Professor of Bioengineering