RT-PCRによるブリからのPhotobacterium damselaの検出

Detection of Photobacterium damsela from Yellowtail by RT-PCR

Matsuoka Satoru

Hirose Keiko

Soumyo Yoshie

Nishizawa Toyohiko

Muroga Kiyokuni

生物生産学研究

Journal of the Faculty of Applied Biological Science, Hiroshima University

魚類病原細菌 Photobacterium damsela subsp. damsela および Ph. damsela subsp. piscicida を検出するために、本菌の 16S rRNA における種特異的塩基配列の解析に基づきRT-PCR法を考案した。まず、魚介類病原菌を中心とした12種のビブリオ属細菌を用いて本RT-PCRの特異性を確認し、また検査魚の脾臓磨砕液をトリプトソーヤ・ブロスで前培養後RT-PCRに供することにより検出感度が高まることが分かった。この方法を用いることにより、外見的に健康な養殖ブリ Seriola quinqueradiata およびウマヅラハギ Thamnaconus modestus やマアジ Trachurus japonicus など数種の天然魚に Ph. damsela が存在することが明らかにされた。

An RT-PCR detection technique was developed for Pholobacterium damsela (=Photobacterium damsela subsp. damsela and Ph. damsela subsp. piscicida) from apparently healthy yellowtail (Seriola quinqueradiata) and other wild fishes, based on the analysis of a species-specific sequence in 16S rRNA of Ph. damsela. Specificity of the RT-PCR with K-F1 (sense primer) and R-GEN (antisense primer) was confirmed by using Ph. damsela subsp. damsela and Ph. damsela subsp. piscicida, 12 strains of fish—associated Vibrio species. Sensitivity of the RT-PCR was improved by combining a pre-culture of the sample (spleen homogenate) with tryptic soy broth (NaCl 2%). This RT-PCR revealed the presence of Ph. damsela from wild yellowtail, filefish (Thamnaconus modestus) and horse mackerel (Trachurus japonicas) caught in the sea as well as apparently healthy cultured yellowtail.

Photobacterium damsela subsp. damsela

Photobacterium damsela subsp. piscicida

Pasteurella piscicida

PCR

rRNA

carrier

Medical sciences [ 490 ]

jpn

広島大学生物生産学部

農林水産研究情報センター

[ISSN] 1341-691X

[NCID] AN10409040