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Identification of South American Leishmania parasite-species using non-radioisoopic DNA probes

Research Project

Project/Area Number 03670204
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 InstitutionKurume University

Principal Investigator

ESHITA Yuki  Kurume University Schooi of Medicine Assistant Professor, 医学部, 講師 (10082223)

Co-Investigator(Kenkyū-buntansha) FUKUMA Toshihide  Kurume University Schooi of Medicine Professor, 医学部, 教授 (90125146)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
KeywordsLeishmania parasite / Sandfly / Southern blot hybridization / Oligonucleotide primer / Leishmania mexicana / Leishmania braziliensis / Non-radioactive probe / Polymerase chain reaction (PCR) / polymerase chain reaction(PCR) / リーシュマニア症 / polymerase chain reaction / プライマー / 非放射性プローブ / リ-シュマニア原虫 / 非放射性プロ-ブ / DNAプロ-ブ
Research Abstract

Three sets synthesized-oligonucleotide primers have been tested for the polymerase chain reaction (PCR). The first set of the primers amplified only the DNA of L.braziliensis complex (L.braziliensis, L.panamensis and L.guyanensis), and the second one amplified only L.mexicana complex (L.amazonensis, L.pifanoi, L.garnhami) except L.mexicana, and the third one amplified L.mexicana as well as all other Leshimania parasite DNA of WHO reference strains. Three sets of them were applied for already identified-Ecuadorian Leishmania parasites by zymodeme analysis. Of fifty-nine of unidentified-Ecuadorian isolates, fifty were identified as L.braziliensis complex parasites by the first set, two isolates were identified as L.mexicana by the second and third ones, and the other five seemed to be other parasites, Trypanosoma sp. by the first, second and third ones of the primers as well as morphological features. Less than 10 parasites were enough numbers to amplify the specific DNA products by PCR.Southern blot hybridization using non-radioactive digoxigenin-DNA probes supported the identification of Leishmania parasites in molecular level. Lutzomyia ayacuhensis specific DNA probe was developed and constructionof the primers is in progress. These results indicate that the PCR using the three sets of the primers may differentiate L.braziliensis complex and L.mexicana complex and L.mexicana from others, especially Trypanosoma parasites, and apply for epidemiological survey of leishmaniasis in South America.

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (35 results)

All Other

All Publications (35 results)

  • [Publications] Urakawa,T.: "Expression of Trypanosoma congolense antigens in Spodoptera frugiperda insect cells" Mol.Biochem.Parasitol.(in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita,Y.: "Detection of Leishmania parasites by DNA amplification using polymerase chain reaction(PCR)" Studies on New World leishmaniasis and its transmission,with particular reference to Ecuador,Research Report Series No.3(Y.Hashiguchi,ed),Kyowa Printing & Co.Ltd.,Kochi,Japan. 16-21 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita,Y.: "Biting activity and Leishmania infection of man-biting species of sandflies,Lutzomyia spp" Studies on New World leishmaniasis and its transmission,with particular reference to Ecuador,Research Report Series No.3(Y.Hashiguchi,ed),Kyowa Printing & Co.Ltd.,Kochi,Japan. 22-27 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita,Y.: "Metacyclic form-specific variable surface glycoprotein-encoding genes of Trypanosoma(Nannomonas)congolense" Gene. 113. 139-148 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 江下優樹: "DNAプローブを用いるトリパノソーマ感染症の診断-非放射性プローブとPCR法-" 日本臨床. 50. 480-485 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita,Y.: "Strategies for the identification of Lutzomyia sandflies and infecting Leishmania parasites,using specific DNA probes for future epidemiologic survey using a molecular biologic techniquein Ecuador and Paraguay" Studies on New World leishmaniasisand its transmission,with particular reference to Ecuador,Research Report Series No.2(Y.Hashiguchi,ed)Kyowa Printing & Co.Ltd.,Kochi,Japan,. 193-207 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 江下優樹: "先進国で得た研究体験を、発展途上国での国際協力に、いかに活かすか.動物・昆虫性疾患と国際交流-衛生動物学の現在・未来-" 神戸、祥文社, 198 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Urakawa, T: "Expression of Trypanosoma congolense antigens in Spodoptera frugiperda insect cells" Mol.Biochem.Parasitol.(in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita, Y: "Detection of Leishmania parasites by DNA amplification using polymerase chain reaction (PCR)" Studies on New World leishmaniasis and its transmission, with particular reference to Ecuador, Research Report Series No.3 (Y.Hashiguchi, ed), Kyowa Printing & Co.Ltd., Kochi, Japan. 16-21 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita, Y: "Biting activity and Leishmania infection of manbiting species of sandflies, Lutzomyia spp" Studies on New World leishmaniasis and its transmission, with particular reference to Ecuador, Research Report Series No.3 (Y.Hashiguchi, ed), Kyowa Printing & Co.Ltd., Kochi, Japan. 22-27 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita, Y: "Metacyclic form-specific variable surface glycoprotein-encoding genes of Trypanosoma (Nannomonas) congolense." Gene. 113. 139-148 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita, Y: "Diagnosis of trypanosomiasis with the parasite specific DNA probes - Non-radioactive probes and PCR methods -" Nippon rinsho. 50 (Spetial edition). 480-485 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita, Y: "Strategies for the identification of Lutzomyia sandflies and infecting Leishmania parasites, using specific DNA probes for future epidemiologicsurvey using a molecular biologic techniquein Ecuador and Paraguay." Studies on New World leishmaniasisand its transmission, with particular reference to Ecuador, Research Report Series No.2 (Y.Hashiguchi, ed), Kyowa Printing & Co.Ltd., Kochi, Japan. 193-207 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Eshita, Y: "How to apply our research experience in developed countries to international cooperation in developing countries Animal-and insect-borne diseases and International exchange - present status and future in Medical zoology -" Shoubun-sha, Kobe Matstumura and Ishii (Editors). 198(123-129) (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Urakawa,T.: "Expression of Trypanosoma congolense antigens in Spodoptera frugiperda insect cells" Mol.Biochem.Parasitol.in press. (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Alexander,J.B.: "New records of phlebotomine sand flies(Diptera:Psychodidae)from Ecuador" Mem.Inst.Oswardo Cruz,Rio de Janeiro. 87. 123-130 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] 江下,優樹: "エクアドル国の高知と低地のリーシュマニア症流行地で採集されたサシチョウバエの卵巣発育について" 衛生動物. 44. 145 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 橋口,義久: "エクアドル高地のアンデス型リーシュマニア症(Andean Ieishmaniasis)" 日熱医会誌. 21. 41 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 江下,優樹: "エクアドル産サシチョウバエから得たpolymerase chain reaction(PCR)産物の性状" 衛生動物. 45. (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 江下,優樹: "polymerase chain Reactionを用いたリーシュマニア原虫のDNA検出(その2)" 寄生虫学誌. 42. (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 江下,優樹: "先進国で得た研究体験を、発展途上国での国際協力に、いかに活かすか 動物・昆虫性疾患と国際交流 -衛生動物学の現在・未来-松村,石井(偏)" 神戸、祥文社, 198 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Eshita,Y.: "Strategies for the identification of Lutzomyia sandflies and infecting Leishmania parasites,using specific DNA probes for future epidemiologic survey using a molecular biologic technique in Ecuador and paraguay." Studies on New World leishmaniasisand its transmission,with particular reference to Ecuador,Research Report Series No.2 (Y.Hashiguchi,ed),Kyowa Printing & Co.Ltd.,Kochi,Japan.

    • Related Report
      1992 Annual Research Report
  • [Publications] Studies on New World leishmaniasis and its transmission,with particular reference to Ecuador,Research Report Series No.3 (Y.Hashiguchi,ed),Kyowa Printing & Co.Ltd.,Kochi,Japan. 193-207 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] Eshita,Y.: "Detection of Leishmania parasites by DNA amplification using polymerase chain reaction (PCR)" Studies on New World leishmaniasis and its transmission,with particular reference to Ecuador,Research Report Series No.3 (Y.Hashiguchi,ed),Kyowa Printing & Co.Ltd.,Kochi,Japan.

    • Related Report
      1992 Annual Research Report
  • [Publications] 16-21 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Eshita,Y.: "Biting activity and Leishmania infection of man-biting species of sandflies,Lutzomyia spp."

    • Related Report
      1992 Annual Research Report
  • [Publications] 22-27 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 江下 優樹: "DNAプローブを用いるトリパノソーマ感染症の診断ー非放射性プローブとPCR法ー" 日本臨床. 50. 480-485 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 江下 優樹: "Polymerase Chain Reactionを用いたリーシュマニア原虫のDNA検出" 日熱医会誌. 19. 70- (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] 橋口 義久: "エクアドル国のリーシュマニア症とその局所療法の試み" 寄生虫学誌. 41. 118- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 江下 優樹,福間 利英: "DNAプロ-ブを用いるトリパノソ-マ感染症の診断ー非放射性プロ-ブとPCR法ー" 日本臨床. 50. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Alexander,J.B.,Eshita,Y.,Labrada,M.Jimenez,M.Gomez,E.A.,Furuya,M.and Hashiguchi,Y.: "Probable transmission of Leishmania panamensis to man by Lutzomyia hartmanni (Diptera:Psychodidae)in Ecuador" (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 江下 優樹,福間 利英,澁谷 敏朗,古谷 正人,橋口 義久: "サシチョウバエの飛来時刻とリ-シュマニア原虫の感染状況" 衛生動物. 42. 190- (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 江下 優樹,福間 利英,山口 晃史,牛島 廣治,澁谷 敏朗,古谷 正人,橋口 義久: "Polymerase Chain Reactionを用いたリ-シュマニア原虫のDNA検出" 日熱医会誌. 19. 70- (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Eshita,Y.,Mimori,T.and Hashiguchi,Y.(Y.Hashiguchi,ed): "Strategies for the identification of Lutzomyia sandflies and infecting Leishmania parasites,using specific DNA probes for future epidemiologic survey using a molecular biologic techniquein Ecuador and Paraguay." Studies on New World leishmaniasisand its tramsmission,with particular ref erence to Ecuador,Research Report Series No.2 Kyowa Printing & Co.Ltd.,Kochi.Japan. 193-207 (1990)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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