Quantification of virus-infected cells using RNA FISH-Flow.
Cody J Warren1, Arturo Barbachano-Guerrero2, Devra Huey1
1Department of Veterinary Biosciences, The Ohio State University, Columbus, OH 43210, USA.
STAR Protocols
|May 20, 2023
Summary
We developed RNA fluorescence in situ hybridization flow cytometry (RNA FISH-Flow) to detect RNA virus infections. This high-throughput method uses fluorescent probes for precise, single-cell analysis of viral dynamics within cell populations.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- RNA viruses pose significant threats to public health.
- Accurate detection of viral infection at the single-cell level is crucial for understanding pathogenesis and developing therapeutics.
Purpose of the Study:
- To present a novel protocol for detecting RNA virus-infected cells.
- To enable high-throughput, single-cell analysis of infection dynamics.
Main Methods:
- Developed RNA fluorescence in situ hybridization flow cytometry (RNA FISH-Flow).
- Utilized 48 fluorescently labeled DNA probes hybridizing to viral RNA (sense or anti-sense).
- Employed flow cytometry for high-throughput population analysis.
Main Results:
- The RNA FISH-Flow protocol enables sensitive detection of viral RNA within cells.
- Probes are adaptable to any RNA virus genome.
- Flow cytometry allows for quantitative, single-cell level assessment of infection.
Conclusions:
- RNA FISH-Flow is a versatile and powerful tool for studying RNA virus infections.
- This method facilitates high-throughput analysis of cellular infection dynamics.
- The protocol provides a foundation for rapid diagnostics and research into viral pathogenesis.
