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抗アラリア剤ピリメサミンの標的酵素遺伝子のクローニングとその薬剤耐性機構の解明

Research Project

Project/Area Number 62570175
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 寄生虫学(含医用動物学)
Research InstitutionOkayama University Medical School

Principal Investigator

ISHII Akira  Dept. Parasitol., Prof., 医学部, 教授 (40012752)

Co-Investigator(Kenkyū-buntansha) WATAYA Yusuke  Fuculty of Pharmaceutical Science. Ass.Prof., 薬学部, 助教授 (90127598)
Project Period (FY) 1987 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1988: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1987: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsMalaria / Pyrimethamine / Drug resistance / ELISA / 倍養 / 培養 / 熱帯熱マラリア / 電気泳動 / OFAGE / 抗マラリア剤
Research Abstract

Malaria parasite, (Plasmodium falciparum) used were FCR-3(Gumma Univ.), FUP(Osaka Univ.) and FCO-1(Okayama Univ.,isolated in North Sumatra) strains. These were adapted to CO2 incubator culture. We tried to make FCR-3 strain resistant to pyrimethamine, methotrexate and trimethoprim by adding increasing doses step by step. They became to show a certain degree of resistance to drugs, however there was a lop of difficulties in attaining high degree of resistance.
We made and assembled an equipment to run pulse field electrophoresis (OFAGE). We run chromosomal large sized DNA of malaria parasites. We could not detect any difference among the strains especially that of gene amplification.
We measured drug susceptibility of the malaria strains. Results were obtained by classical culture method, however it took more than 3 days for us to obtain drug resistance information and needed time consuming microscopic examination. We tried to use anti-BrdU antibody in ELISA system to measure incorporation of bromodeoxyuridine into the parasites. The system was established as a novel short time assay for drug susceptibility testing. The method was proved useful in the field in North Sumatra.
Resistance to pyrimethamine was reported to be caused by a point mutation in dehydrofolate reductase gene. We tried to prepare the probe to detect not only malaria parasite but also mutation of the gene. The fundamental experimentaions are under way.

Report

(3 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • 1987 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] H.Doi;A.Ishii;K.Shimono.: Trans.Roy.Soc.Trop.med.& Hug.82. 190-193 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] H:Doi,Safei;A:Ishii: Japanese Journal of Topical Medicine and rtygiere. (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] H.Doi; A.Ishii; K.Shimono.: "A rapid in vitro assay system using anti-bromodioxyuridine for drug susceptibility of Plasmodium falciparum." Transection of the Royal Society for Tropical Medicine and Hygiene. 82. 190-193 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] H.Doi, Safei; A.Ishii.: "In vitro bromodeoxyuridine incorporation assay for drug susceptibility of Plasmodium falciparum in the field." Japanese Journal of TYropical Medicine and Hygiene. (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] H.Doi,;A.Ishii.;K.Shimono.: Jrans.Rey.Soc.Trop.Med.&Hyg.82. 190-193 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] H. Doi;A. Ishii;K. Shimono: Trans. Roy. Soc. Trop. Med. & Hyg.82. 190-193

    • Related Report
      1987 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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