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Development of new gene diagnosis by detecting NTPase gene of Toxoplasma gondii.

Research Project

Project/Area Number 06557018
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section試験
Research Field 寄生虫学(含医用動物学)
Research InstitutionKeio University, School of Medicine,

Principal Investigator

ASAI Takashi  Keio University, School of Medicine, Department of Tropical Medicine and Parasitology, Assistant Professor, 医学部, 講師 (50175163)

Co-Investigator(Kenkyū-buntansha) UEDA Masakazu  Keio University, School of Medicine, Department of Tropical Medicine and Parasit, 医学部, 講師 (50142419)
OKUZAWA Eiichi  Keio University, School of Medicine, Department of Tropical Medicine and Parasit, 医学部, 助手 (20177166)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥3,000,000 (Direct Cost: ¥3,000,000)
KeywordsParasite / Protozoa / Toxoplasma gondii / NTPase / Diagnosis / RT-PCR / PCR
Research Abstract

We tried to amplify a gene fragment (about 800 base pair) in NTPase, which is present in tachyzoite cells causing acute toxoplasmosis, gene by PCR and RT-PCR using DNA and RNA extracted from tissues of animals infected with Toxoplasma gondii and suspected human patient for toxoplasmosis as the tempelates. The results indicated that it was rigidly possible to detect the gene and mRNA of NTPase from the tissues of animal infected with T.gondii. Then, we tried to detect the gene for diagnosis of a suspected human patient as congenital toxoplasmosis. We succeeded to detect the mRNA but not to detect the DNA suggesting that RT-PCR was more sensitive than simple PCR.Anyway, we succeeded to diagnose congenital toxoplasmosis within a short period. Preliminal experiment indicated that only one tachyzoite cell was detected by RT-PCR and the sensitivity of RT-PCR was 10 times higher than that of PCR.The reason why RT-PCR is more sinsitive than PCR is that tachyzoite cells contain lot of mRNA and copy number of NTPase is abundant. In the present study, we have succeeded to detect NTPase mRNA by RT-PCR indicating that RT-PCR is useful for diagnosis of acute toxoplasmosis in general. Comparing with previous diagnostic methods for acute toxoplasmosis, which took long time to complete, this RT-PCR and PCR methods have a merit being practicable to diagnose acute toxoplasosis at once.

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Jun-ichi Sanuki et al.: "Identification of Entamoeba histolytica and E-disparcysts in stool by polymerase chain reaction" Parasitol.Res. 83. 96-98 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takashi Asai et al.: "Biochemical and Molecular-characterization of -" J.Biol.Chem.270. 11391-11397 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] L.D.Sibley et al.: "Toxoplasma gorolii:Secretion of potent-" Exptl.Parasitol.79. 301-311 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 浅井隆志、他: "ニューモシスチス・カリニおよびトキソプラズマの生物学" カレントテラピー. 14. 160-164 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 浅井隆志、他: "急性トキソプラズマ症の新しい検査法" Biomedical News. 23-11. 3-4 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 佐貫潤一、他: "RT-PCRを用いた先天性トキソプラズマ症の診断法" Clinical Parasitology. 6. 25-27 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Jun-ichi Sanuki, Takashi Asai, Eiichi Okuzawa, Seiki Kobayashi, Tsutomu Takeuchi: "Identification of Entamoeba histolytica and E.dispar cysts in stool by polymeras chain reaction." Parasitol.Res.83. 96-98 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takashi Asai, Satoshi Miura, L.David Sibley, Hironori Okabayashi, Tsutomu Takeuchi: "Biochemical and Molecular Characterization of Nucleoside Triphosphate Hydrolase Isozymes from the Parasitic Protozoan Toxoplasma gondii" J.Biol.Chem.270. 11391-11397 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] L.David Sibley, Ingrid R.Niesman, Takashi Asai, Tsutomu Takeuchi: "Toxoplasma gondii : Secretion of a Potent Nucleoside Triphosphate Hydrolase into the Parasitophorous Vacuole" Exptl.Parasitol. 79. 301-311 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takashi Asai, Tsutomu Takeuchi: "Biology of Pneumocystis carinii and Toxoplasma gondii (in Japanese)" Current Therapy. 14. 160-164 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takashi Asai, Jun-ichi Sanuki: "A new diagnostic technique for acute toxoplasmosis (in Japanese)" Biomedical News. 23-11. 3-4 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Jun-ichi Sanuki, Takashi Asai: "Gene diagnosis of Entamoeba histolytica (in Japanese)" Biomedical News. 23-8. 3-4 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Jun-ichi Sanuki, Takashi Asai, Tsutomu Takeuchi, Others: "Usefulness of RT-PCR for diagnosis of congenital toxoplasmosis (case of patient and technique) (in Japanese)" Clinical Parasitology. 6. 25-27 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Jun-ichi Sanuki, Takashi Asai, Eiichi Okuzawa, Tsutomu Takeuchi, others.: "Detection of Entamoeba histolytica and diagnosis of amebiasis by PCR using endoscopic biopsy sample from large intestine (in Japanese)" Clinical Parasitology. 6. 114-115 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Eiichi Okuzawa, Takashi Asai: "200 Strategies for common diseases, Parasite/Parasitic diseases by protozoa/amebiasis (in Japanese)" Medicina. 32. 544-546 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takashi Asai (Review): "Nucleoside triphosphate hydrolase which is present uniquely in Toxoplasma gondii (in Japanese)" Japan J.Protozoology. 27. 5-10 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Jun-ichi Sanuki et al: "Identification of Entamoeba histolytica and E-dispar cysts in stool by polymerase chain reaction" Parasitol.Res.83. 96-98 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 浅井隆志、左貫潤一: "急性トキソプラズマ症の新しい検査法" Biomedical News. 23-11. 3-4 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 左貫潤一、浅井隆志: "赤痢アメーバ症の遺伝子診断" Biomedical News. 23-8. 3-4 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 浅井隆志、竹内勤: "ニューモシスチス・カリニおよびトキソプラズマの生物学" カレントテラピー. 14-7. 160-164 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Takashi Asai et al: "Biochemical and Molecular Characterization----" Journal of Biological Chemistry. 270. 11391-11397 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 佐貫潤一,浅井隆志,他: "RT-PCRを用いた先天性トキソプラズマ症の診断法の有用性について" Clinical Parasitology. 6. 25-27 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Takashi Asai et al: "Biochemical and Molecular Characterigation・・・・" Journal of Biological Chemistry. 270(印刷中). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] L.D.Sibley et al: "Toxoplasma gondii secretion・・・・・・・・" Experimental Parasitology. 79. 301-311 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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