Colorimetric immunocapture loop mediated isothermal amplification assay for detection of Impatiens necrotic spot virus (INSV) by GineFinder™ dye

Abstract

Impatiens necrotic spot virus (INSV) has been seen in many types of ornamental plants, including annuals and perennials. The virus is rapidly spread by the insect vector, the western flower thrips, and there is no cure for the disease. Due to its potential spread to other plants grown in the same area, it is very important to act quickly to remove and destroy affected plants. Therefore, a rapid diagnosis of INSV is crucial so that growers can determine if plants are diseased and act without delay. Loop-mediated isothermal amplification (LAMP) or reverse transcription LAMP (RT-LAMP) assay is a technique for amplifying DNA or RNA under a constant temperature, with high specificity, sensitivity, rapidity and efficiency. We developed a colorimetric immunocapture reverse transcription (IC-RT)-LAMP assay to detect INSV using RT-LAMP and a nucleic acid dye, GeneFinder™. We accordingly propose this assay as a highly reliable alternative viral recognition system for INSV detection.