Rapid Detection of RSV, RBSDV, and SRBSDV in Plants and Insect Vectors by the RT-RPA-LF Assay

Area of Science:

• Plant pathology
• Virology
• Molecular diagnostics

Background:

• Accurate detection of plant viruses is crucial for disease management.
• Rice viruses like RSV, RBSDV, and SRBSDV cause significant crop losses.
• Existing detection methods can be time-consuming and require laboratory facilities.

Purpose of the Study:

• To develop a rapid, on-site detection method for RSV, RBSDV, and SRBSDV.
• To enable simultaneous detection in both plant tissues and insect vectors.
• To facilitate early diagnosis and disease management.

Main Methods:

• Developed a reverse transcription recombinase polymerase amplification combined with lateral flow strip (RT-RPA-LF) assay.
• Designed virus-specific primers targeting conserved genomic regions.
• Integrated the assay with a paper-based RNA rapid extraction method for field use.

Main Results:

• The RT-RPA-LF assay demonstrated high specificity with no cross-reactivity.
• Sensitivity limits were 10^9-fold dilution for SRBSDV and 10^8-fold dilution for RSV/RBSDV in plants.
• The assay could detect a single viruliferous planthopper among 1000 individuals.
• Field detection accuracy was comparable to conventional RT-PCR.

Conclusions:

• The RT-RPA-LF method provides rapid, simple, specific, and visualizable on-site detection of rice viruses.
• This assay is effective for both plant and insect vector samples.
• The developed method facilitates early diagnosis and timely disease management strategies.