| Project/Area Number |
06454095
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General fisheries
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
YOSHIMIZU Mamor Hokkaido Univ., Fac.Fish., Associate Prof., 水産学部, 助教授 (40122915)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIZAWA Toyohiko Hiroshima Univ., Fac.of Appl.Biol.Sci., Asist.Prof., 生物生産学部, 助手 (10222184)
SAWABE Tomoo Hokkaido Univ., Fac.of Fish., Assist.Prof., 水産学部, 助手 (30241376)
TAJIMA Kenichi Hokkaido Univ., Fac.of Fish., Associate Prof., 水産学部, 助教授 (80002252)
EZURA Yoshio Hokkaido Univ., Fac.of Fish., Prof., 水産学部, 教授 (80001618)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 1996: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1995: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1994: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | PCR / viral nervous necrosis / VNN / infectious hematopoietic necrosis / IHN / IPN / ELISA / ウイルス性神経壊死症 / IHNV / IPNV / HRV / 神経壊死症 / サケ科魚類 / マツカワ / ヒラメ / 魚類培養細胞 / プライマー / ビオチン化プローブ / RTE-2 |
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| Research Abstract |
Viral disease is a serious problems of fish in hatchery. Especially, viral nervous necrosis of marine fish and infectious hematopoietic necrosis of salmonids are difficult to control. In this study, we studied and established the highly sensitive and specific diagnostic method to detect the virus specific gene using polymerase chain reaction (PCR) method. Results obtained in this study are summarized as follow.
1. Striped jack nervous necrosis virus (SJNNV) coat protein gene and infectious hematopoietic necrosis virus (IHNV) N protein gene were amplified by polymerase chain reaction (PCR) an could apply for the specific and sensitive diagnosis of VNN and IHN.
2. Rainbow trout embryo (RTE-2) and sea bass kidney (SBK-2) cell lines were selected for the isolation of IHNV and NNV.
3. Method of the simultaneous detection and identification of both IHNV and infectious pancreatic necrosis virus (IPNV) in a single PCR reaction was developed.
4. Fish nodaviruses were clasified into 4 clusters by molecular phylo-genetic analysis of the coat protein gene.
5. Combination of RT-PCR and antibody screening by ELISA was effective method for the identification of nervous necrosis viurs carriers in a flounder brood stock.
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