Rapid visual detection of the severely destructive invasive pest Carpomya pardalina: a novel loop-mediated isothermal amplification (LAMP) assay for efficient on-site diagnostics.

Carpomya pardalina, primarily found in certain regions of Eastern Europe, Central Asia and North Africa, is a highly serious pest that spreads rapidly. Its host plants have an extremely wide range, rendering C. pardalina highly vulnerable to long-distance spread through the import and export of agricultural products. Its invasion could have a devastating impact on relevant industries. Given the difficulty in differentiating Tephritidae pests based on morphological characteristics, a rapid and accurate molecular identification method is required to shorten the identification period of C. pardalina. We have developed a loop-mediated isothermal amplification (LAMP) assay for C. pardalina. A set of LAMP primers was designed for the COX1 gene of C. pardalina. The LAMP amplification was capable of distinguishing C. pardalina from its similar species with excellent specificity, and the reaction demonstrated remarkable sensitivity, enabling the detection of a DNA template at a concentration of 0.969 ng/μL. Moreover, the amplification could be performed swiftly using the DNA extracted from crude DNA extractions as a template. The detection can be completed within 75 min from DNA extraction, significantly enhancing the detection speed. Meanwhile, the visualization test demands simple equipment and is easy to operate, making it suitable for use at the port front line and under field working conditions. In this study, a set of rapid and accurate techniques was established based on the loop-mediated isothermal amplification (LAMP) technology for the detection of C. pardalina. © 2025 Society of Chemical Industry.