# Rapid detection of Impatiens necrotic spot virus from thrips vectors using reverse transcription-recombinase polymerase amplification

## TL;DR

A new rapid test detects a plant virus in thrips, helping predict and manage outbreaks in agriculture.

## Contribution

A cost-effective RT-RPA assay for rapid INSV detection in thrips vectors is developed.

## Key findings

- The RT-RPA assay detects INSV in 25 minutes at a constant temperature of 42°C.
- The method successfully identified and quantified INSV in both larval and adult thrips from field populations.
- The assay offers sensitivity comparable to existing detection methods but with faster results.

## Abstract

The plant virus, Impatiens necrotic spot virus (INSV), is an economically important pathogen of vegetables, fruits, and ornamental crops. INSV is vectored by the western flower thrips, Frankliniella occidentalis, a small insect pest that is globally distributed. In recent years, INSV outbreaks have reached epidemic levels in the Salinas Valley of California—an agriculturally rich region where most of the lettuce (Lactuca sativa) is produced in the United States. Due to the obligate nature in which virus transmission occurs, new tools that could rapidly detect INSV from thrips vectors would enhance our ability to predict where virus outbreaks may occur. Here, we report on the development of a reverse transcription-recombinase polymerase amplification (RT-RPA) assay that can detect INSV from individual thrips. The assay uses crude extraction methods, is performed at a single temperature of 42 °C, can be completed in 25 min, and provides sensitivity levels that are comparable to other available detection methods. When the assay was used on field populations of thrips, INSV was successfully identified and quantified from individual larvae and adults. The work provides a new cost-effective surveillance tool that can rapidly detect INSV from its insect vector and from plants.

The online version contains supplementary material available at 10.1038/s41598-024-73078-4.