Real-time PCR assay for detection of Environmental DNA from Chironex yamaguchii

• Published in: Medical Entomology and Zoology Vol. 74; no. 1; pp. 13 - 20
• Main Authors: Azama, Yasuhito, Oka, Shin-ichiro, Teruya, Morimi, Toshino, Sho, Tanimoto, Miyako, Hanahara, Nozomi, Kuba, Yumani, Miyagi, Ayano, Fukuchi, Yoshimune
• Format: Journal Article
• Language: English; Japanese
• Published: Tokyo The Japan Society of Medical Entomology and Zoology 25.03.2023; Japan Science and Technology Agency
• Subjects: Chironex yamaguchii; Cubozoa; Cytochrome-c oxidase; envenomation; Environmental DNA; Genetic testing; Polymerase chain reaction; species-specific real-time PCR assay; Venom; Japan; Okinawa
• ISSN: 0424-7086; 2185-5609
• DOI: 10.7601/mez.74.13

The box jellyfish Chironex yamaguchii is a highly venomous cubomedusa, also known as “habu-kurage” in Japan. It is a significant threat to marine activities in Okinawa in southern Japan. Annually, envenomation of the species causes nearly 100 to 200 injuries in the Okinawa Prefecture. The present study developed a species-specific real-time PCR assay to analyze environmental DNA (eDNA) for rapid detection of jellyfish in water samples. Primers and probe were designed based on the sequences of the mitochondrial cytochrome c oxidase subunit I region using tissue samples of nine specimens obtained from the Okinawa Prefecture. The specificity of the real-time PCR assay was confirmed by amplifying the DNA samples of 10 different box jellyfish species. A practical assay for eDNA detection was successful on water samples obtained from the natural habitat of C. yamaguchii. In addition, the degradation rate of eDNA in the tank experiment was 65% of the initial concentration at 1 h, 22% at 12 h, and 3% at 48 h.