1995 Fiscal Year Final Research Report Summary
Vibriosis of kuruma prawn : Detection of the causative bacterium by PCR
| Project/Area Number |
06660238
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General fisheries
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
MUROGA Kiyokuni Hiroshima Univ.Fac.Appl.Biol.Sci., Professor, 生物生産学部, 教授 (30011993)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAI Toshihiro Hiroshima Univ.Fac.Appl.Biol.Sci., Associate Professor, 生物生産学部, 助教授 (60164117)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Vibrio penaeicida / PCR (RT-PCR) / Kuruma prawn / Vibriosis / detection / rRNA / primer |
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| Research Abstract |
Taxonomical position of the causative bacterium of vibriosis in kuruma prawn, Vibrio sp.PJ was established and a detection methood for the pathogen by PCR method was developed.
1. Vibrio penaeicida n.sp.was proposed as the name of the pathogen.
2. It was found that the pathogen multiplied in the stomach, midgut gland and lymphoid organs of experimentally infected kuruma prawns, but a detection method with higher sensitivity was needed for the determination of the initial multiplication site.
3. Based on the base sequence data of Vibrio 16SrRNA,species-specific sequence was determined by cloning.
4. Based on the above specific sequence, a set of primers was designed for RT-PCR.The aimed product was obtained from all strains of V.penaeicida but not from other vibrios.
5.It was found that GITC-chlorophorm or Tween20-phenol method was applicable for the extraction of RNA from kuruma prawn.
6. The above RT-PCR method gave higher detectin rate of V.penaeicida from carrier kuruma prawns than conventional culture method.
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