Liu J, An T, Peng J, Zhu Q, Zhao H, Liang Z, Mo K,. An amplification-free digital droplet assay for influenza A viral RNA based on CRISPR/Cas13a. Analyst. 2025 Feb 14
Most of the CRISPR-based RNA detection methods are combined with amplification to improve sensitivity, which lead to some drawbacks such as aerosol pollution, complicated operation, and amplification bias. To address the above issues, we developed a digital detection method for influenza A viral RNA based on droplet microfluidics and CRISPR/Cas13a without polymerase chain reaction. We used a microsphere coupled to a capture probe to extract and concentrate the target RNA from the samples, and then restricted the target-induced CRISPR/Cas13a cleavage event to microfluidic droplets, thus enhancing the local signal intensity and enabling single-molecule detection. With a detection limit of 10 copies per μL, influenza A viral RNA can be detected in less than 1 h. Both clinical and synthetic series samples were used to validate the assay´s performance. With the help of this direct RNA diagnostic method, a variety of RNA molecules can be easily and accurately detected at the single-molecule level. This research has broad prospects in clinical applications.
See Also:
Latest articles in those days:
- Generation of Nasal Cell-Derived Human Alveolar Organoids and Organoid-Macrophage Assembloids for in Vitro Lung Modeling 6 hours ago
- Determinants of the Seasonal Influenza Vaccination Uptake Among People Aged 50 Years or Above During and After the Pandemic: A Systematic Review 18 hours ago
- [preprint]A GIS-based framework for standardized environmental characterization in One-Health surveillance: a case study of HPAI monitoring in wetlands 18 hours ago
- First detection and transatlantic introduction of Influenza A(H3N2) subclade K (J.2.4.1) into Ecuador: insights from genomic sentinel surveillance 18 hours ago
- Assessment of influenza virus and coronavirus tropism, replication competence and disease severity in ex vivo and in vitro cultures of the human respiratory tract 2 days ago
[Go Top] [Close Window]


