Publisher : The Korean Society of Fisheries and Aquatic Science
DOI : 10.5657/KFAS.2015.0731
Title & Authors
Molecular Identification and Development of a PCR Assay for the Detection of a Philometrid Nematode in Rockfish Sebastes schlegeli Seo, Han-Gill; Seo, Jung Soo; Ryu, Min Kyung; Lee, Eun Hye; Jung, Sung Hee; Han, Hyun-Ja;
Nematode infection in the epithelial tissue of cultured rockfish Sebastes schlegeli was first reported in 2012. Since then, nematode infections have caused serious economic losses in rockfish aquaculture on the west coast of Korea. Taxonomic and life cycle information for this parasite are currently unknown. In this study, 18S rRNA and cytochrome c oxidase subunit I (COI) genes were used for molecular identification and polymerase chain reaction (PCR) to detect the invisible stages of this parasite. Nucleotide sequences of the 18S rRNA of the rockfish nematode showed 98% identity with that of Philometra morii. Therefore, this rockfish nematode was classified to the Philometridae family. However, we could not identify it to genus level using 18S rRNA. Its COI nucleotide sequences shared 85% and 82% identities with those of Bursaphelenchus sinensis and Philometra overstreeti, respectively. In addition, two gene-specific primer sets were designed based on the 18S rRNA gene to detect the intermediate host and nematode larvae. These primers were specific to this rockfish nematode without cross-reacting to other pathogens. The detection limit of the PCR assay using these primers was 1,000 copies of nematoda plasmid DNA. Therefore, the PCR assay described here is suitable for the detection of nematode DNA within rockfish. In addition, this PCR assay could be used to detect nematode larvae and the intermediate host.
Philometrid nematode;Rockfish;18S rRNA gene;Cytochrome c oxidase subunit I gene;PCR;