Location: Crop Improvement and Protection Research
Title: Rapid real-time detection of Impatiens necrotic spot virus from western flower thrips using reverse transcription-recombinase polymerase amplificationAuthor
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Submitted to: American Phytopathological Society Annual Meeting
Publication Type: Abstract Only Publication Acceptance Date: 6/5/2024 Publication Date: N/A Citation: N/A Interpretive Summary: Technical Abstract: Impatiens necrotic spot virus (INSV; genus Orthotospovirus, family Tospoviridae) is a negative-sense single-stranded RNA virus with a tripartite genome that infects over 650 plant species, including economically important species such as lettuce and tomato. The virus is transmitted in a persistent manner, primarily by the insect western flower thrips (WFT) (Frankliniella occidentalis). Infection of lettuce by INSV causes necrotic leaf spots, stunted plant growth, and plant death. The emergence and re-emergence of INSV in lettuce has recently occurred in several places in the United States, including California and Arizona, resulting in serious economic losses to the industry. The availability of rapid and accurate tools for detecting INSV from WFT would be important for identifying vector populations that are associated with the virus. In this report, we describe the development of a new molecular assay for rapid real-time detection of INSV from thrips and lettuce. The assay utilizes isothermal technology of reverse transcription-recombinase polymerase amplification (RT-RPA) and the entire process from sample to result can be completed within 25 min. All reactions are performed at a constant temperature in a single PCR tube. The assay specifically detects INSV at a minimum concentration of 1 fg of target RNA. The assay was also shown to be as sensitive as RT-qPCR and more sensitive than RT-PCR, TAS-ELISA, and Immunostrip assays. To our knowledge, this is the first application of RT-RPA for the detection of INSV from thrips and lettuce. |
