Abstract

Invasive alien species pose significant threats to global biodiversity, and early detection is critical for effective management. Cryptotermes domesticus is a destructive pest known to infest timber, furniture, and wooden structures, causing economic and ecological damage. This study aimed to develop a high-specificity and highly- sensitive quantitative polymerase chain reaction (qPCR) probe-based assay for precise identification of C. domesticus. C. domesticus samples, collected from an apartment in the Seoul metropolitan area, South Korea, specifically from the interior of a sliding door frame infested with termites, were subjected to whole-genome sequencing to pinpoint species-specific genetic sequences. Species-specific primer sets were designed based on whole-genome sequencing data, and after the evaluation of the amplification efficiency and specificity through qPCR, the optimal primer set was selected. The developed qPCR assay effectively distinguished the target species without cross-reactivity with native termite species or foreign termite species confirmed to have entered the country. The molecular diagnostic method developed in this study enables quick and accurate detection of C. domesticus, offering a reliable tool with potential applications in pest management and ecological research.