Spodoptera eridania and S. ornithogalli (Lepidoptera: Noctuidae), which are polyphagous pests that damage various crops such as tomatoes and beans are regulated quarantine species that are highly likely to invade South Korea. Therefore, it is crucial to promptly and accurately identify the presence of S. eridania and S. ornithogalli in crop fields to effectively eradicate as a regulated quarantine species. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay, which allows for rapid in-field identification. To develop the LAMP　assay, we selected target species-specific genomic regions from the whole-genome sequences of one target and 13 other lepidopteran species. We validated each five and six primer sets that consistently produced positive reactions in S. eridania and S. ornithogalli, respectively. To test the sensitivity of the each locus, LAMP reactions were performed using various reaction times using crude DNA, which was extracted from various types of adult tissues. All sensitivity tests were also successful.

Loop-mediated isothermal amplification (LAMP) is a rapid, specific, cost-effective detection method by amplifying nucleic acid under isothermal conditions. In this study, we used LAMP for detection of Hamiltonella defensa that lives as a facultive endosymbiont of whitefly ‘Bemisia tabaci’. We designed the Hamiltonella-specific primers by targeting 16S ribosomal RNA gene and validated the specificity of one primer set. To find the optimum temperature for our primer set, the LAMP reaction was held at the temperature, 60℃, 62℃ and 65℃. As a result, 62℃ was the optimum reation temperature for LAMP reaction. Specificity of primer set was tested by the reaction to both Trialeurodes vaporariorum and B. tabaci. After the whole procedure, the amplicons by LAMP were visualized by adding SYBR Green to the reaction tube.