{"created":"2023-05-15T15:17:00.304619+00:00","id":3004,"links":{},"metadata":{"_buckets":{"deposit":"8d647c36-073f-4008-9f1a-6af79f2acbe6"},"_deposit":{"created_by":12,"id":"3004","owners":[12],"pid":{"revision_id":0,"type":"depid","value":"3004"},"status":"published"},"_oai":{"id":"oai:obihiro.repo.nii.ac.jp:00003004","sets":["242:243"]},"author_link":["164","63","295","340","5360","312"],"item_6_alternative_title_1":{"attribute_name":"その他（別言語等）の研究課題名","attribute_value_mlt":[{"subitem_alternative_title":"Development of Diagnostic Methods for African Trypanosomes and Their Evaluation Using Field Samples","subitem_alternative_title_language":"en"}]},"item_6_contributor_5":{"attribute_name":"研究分担者","attribute_value_mlt":[{"contributorAffiliations":[{"contributorAffiliationNameIdentifiers":[{"contributorAffiliationScheme":"ISNI","contributorAffiliationURI":"http://www.isni.org/isni/"}],"contributorAffiliationNames":[{"contributorAffiliationNameLang":"ja"}]}],"contributorNames":[{"contributorName":"Xuan, Xuenan","lang":"en"},{"contributorName":"玄, 学南","lang":"ja"}],"familyNames":[{},{}],"givenNames":[{},{}],"nameIdentifiers":[{"nameIdentifier":"295","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"9000002841347","nameIdentifierScheme":"CiNii ID","nameIdentifierURI":"http://ci.nii.ac.jp/nrid/9000002841347"},{"nameIdentifier":"10292096","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=10292096"},{"nameIdentifier":"read0055834","nameIdentifierScheme":"researchmap","nameIdentifierURI":"https://researchmap.jp/read0055834"}]},{"contributorNames":[{"contributorName":"田仲, 哲也","lang":"ja"}],"nameIdentifiers":[{"nameIdentifier":"5360","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"00322842","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=00322842"}]},{"contributorAffiliations":[{"contributorAffiliationNameIdentifiers":[{"contributorAffiliationScheme":"ISNI","contributorAffiliationURI":"http://www.isni.org/isni/"}],"contributorAffiliationNames":[{"contributorAffiliationNameLang":"ja"}]}],"contributorNames":[{"contributorName":"Yokoyama, Naoaki","lang":"en"},{"contributorName":"横山, 直明","lang":"ja"}],"familyNames":[{},{}],"givenNames":[{},{}],"nameIdentifiers":[{"nameIdentifier":"312","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"9000002627114","nameIdentifierScheme":"CiNii ID","nameIdentifierURI":"http://ci.nii.ac.jp/nrid/9000002627114"},{"nameIdentifier":"80301802","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=80301802"},{"nameIdentifier":"read0074761","nameIdentifierScheme":"researchmap","nameIdentifierURI":"https://researchmap.jp/read0074761"}]},{"contributorAffiliations":[{"contributorAffiliationNameIdentifiers":[{"contributorAffiliationNameIdentifier":"","contributorAffiliationScheme":"ISNI","contributorAffiliationURI":"http://www.isni.org/isni/"}],"contributorAffiliationNames":[{"contributorAffiliationName":"","contributorAffiliationNameLang":"ja"}]}],"contributorNames":[{"contributorName":"Nagasawa, Hideyuki","lang":"en"},{"contributorName":"長澤, 秀行","lang":"ja"}],"familyNames":[{},{}],"givenNames":[{},{}],"nameIdentifiers":[{"nameIdentifier":"164","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"60172524","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=60172524"},{"nameIdentifier":"read0017304","nameIdentifierScheme":"researchmap","nameIdentifierURI":"https://researchmap.jp/read0017304"}]},{"contributorNames":[{"contributorName":"藤崎, 幸蔵","lang":"ja"}],"nameIdentifiers":[{"nameIdentifier":"340","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"9000006874417","nameIdentifierScheme":"CiNii ID","nameIdentifierURI":"http://ci.nii.ac.jp/nrid/9000006874417"},{"nameIdentifier":"00292095","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=00292095"}]}]},"item_6_description_10":{"attribute_name":"研究代表者番号","attribute_value_mlt":[{"subitem_description":"10271751","subitem_description_type":"Other"}]},"item_6_description_11":{"attribute_name":"研究機関","attribute_value_mlt":[{"subitem_description":"帯広畜産大学","subitem_description_language":"ja","subitem_description_type":"Other"}]},"item_6_description_13":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"LAMP法によるトリパノソーマ特異的遺伝子検出法の開発。\nLAMP法は等温反応で高感度遺伝子増幅を行う方法である。また、遺伝子増幅の有無が反応液の濁度によって判別可能である。従って実施には単純なインキュベーター以外の特別な設備を要しない。LAMP法のこのような特徴は、十分な権設・設備が利用できない開発途上国や小規模診療施設などで高感度病原体検出を実施する上で非常に有用である。トリパノソーマ慢性感染マウスを用いた実験では、直接検出<PCR<LAMPの順に検出感度が高くなり、LAMP法の有用性が検証された。従来から指摘されていた通り本実験においても、LAMP法に用いるBst DNAポリメラーゼはPCR法に使用するTaq DNAポリメラーゼよりも血液由来酵素阻害因子の影響を受けにくいことを示唆する結果が得られた。タイ国・農務省・農産物食品局(National Bureau of Agricultural Commodity and Food Standards(ACFS), Ministry of Agriculture and Cooperatives, Bangkok, Thailand)のツンタスヴァン(D.Tuntasuvan)博士との共同研究で、トリパノソーマ感染ブタからのトリパノソーマ検出を、直接検出法(スメア法、遠心法)、マウス接種法、PCR法、LAMP法の各方法を用いて行った。その結果、LAMP法はPCR法と同等以上の高感度でトリパノソーマ遺伝子を検出した。反応所要時間および検出の簡便性ではLAMP法がPCR法よりも優れている。今回の研究で、LAMP法の野外使用における有用性は十分証明されたと考えるが、今後LAMP法に供するDNAサンプルの調製法については十分検討すべきである。LAMP法は高感度であるため、今回の研究においてもDNA調製段階での試料間の相互汚染が問題となることがあった。\n血清診断用抗原の探索\nトリパノソーマミトコンドリアHSP70およびリボソームP0蛋白質がアフリカトリパノソーマ検出ELISA抗原として有用であることが明らかとなった。リボソームP0蛋白質はC末端に強い抗原性を示す領域を有することも明らかとなった。P0-C末端領域の合成ペプチドを用いたELISAでもトリパノソーマ感染血清を良好に検出可能であった。合成ペプチドを使用することによって組換え蛋白質の発現と精製など大幅に診断系構築のステップを削減することが可能となる。","subitem_description_language":"ja","subitem_description_type":"Abstract"},{"subitem_description":"Loop-mediated isothermal amplification of DMA (LAMP)\nThe loop-mediated isothermal amplification (LAMP) reaction is a method that amplifies DNMA with high specificity, efficiency, and rapidity under isothermal conditions with only simple incubators. An added advantage of LAMP over PCR-based methods is that DNA amplification can be monitored spectrophotometrically and/or with the naked eye without the use of dyes. Here we report our conditions for a highly sensitive, specific, and easy diagnostic assay based on LAMP technology for the detection of parasites in the Trypanosoma brucei group (including T.brucei brucei, T.brucei gambiense, T.brucei rhodesiense, and T.evansi) and T.congolense. We show that the sensitivity of the LAMP-based method for detection of trypanosomes in vitro is up to 100 times higher than that of PCR-based methods. In vivo studies in mice infected with human-infective T.brucei gambiense further highlight the potential clinical importance of LAMP as a diagnostic tool for the identification of African trypanosomiasis.\nELISA\nThe ability to use mitochondrial heat shock protein 70 (MTP) of Trypanosoma congolense as a diagnostic antigen was examined. The B-cell epitope recognized by MAb 10F9 was located within 206 amino acids from the C terminus. Depending on the conditions of protein extraction, MTP was cleaved into smaller polypeptides by endogenous proteases. However, the C-termmal epitope of MTP was preserved with a high degree of antigenicity, even after cleavage. Antibody detection by enzyme-linked immunosorbent assay with the truncated recombinant MTP revealed that anti-MTP antibodies exist in experimentally infected mouse sera. Thus, MTP may be useful as an antigen for the serodiagnosis of primary T.congolense infection.","subitem_description_language":"en","subitem_description_type":"Abstract"}]},"item_6_description_14":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"平成13年度～平成15年度科学研究費補助金基盤研究(B)(2)研究成果報告書","subitem_description_language":"ja","subitem_description_type":"Other"}]},"item_6_description_19":{"attribute_name":"フォーマット","attribute_value_mlt":[{"subitem_description":"application/pdf","subitem_description_type":"Other"}]},"item_6_description_9":{"attribute_name":"研究課題番号","attribute_value_mlt":[{"subitem_description":"13556048","subitem_description_type":"Other"}]},"item_6_text_12":{"attribute_name":"助成元","attribute_value_mlt":[{"subitem_text_language":"ja","subitem_text_value":"科学研究費助成事業"}]},"item_6_version_type_20":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2018-01-30"}],"displaytype":"detail","filename":"4827.pdf","filesize":[{"value":"6.6 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"4827.pdf","url":"https://obihiro.repo.nii.ac.jp/record/3004/files/4827.pdf"},"version_id":"2c73e3de-3053-4744-b0c7-0667521e7dfc"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_researcher":{"attribute_name":"研究代表者","attribute_type":"creator","attribute_value_mlt":[{"creatorAffiliations":[{"affiliationNameIdentifiers":[{"affiliationNameIdentifierScheme":"ISNI","affiliationNameIdentifierURI":"http://www.isni.org/isni/"}],"affiliationNames":[{"affiliationNameLang":"ja"}]}],"creatorNames":[{"creatorName":"Inoue, Noboru","creatorNameLang":"en"},{"creatorName":"井上, 昇","creatorNameLang":"ja"}],"familyNames":[{},{}],"givenNames":[{},{}],"nameIdentifiers":[{},{},{},{}]}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"トリパノソーマ病特異的診断法の開発および野外材料を用いた実用評価","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"トリパノソーマ病特異的診断法の開発および野外材料を用いた実用評価","subitem_title_language":"ja"}]},"item_type_id":"6","owner":"12","path":["243"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2009-03-03"},"publish_date":"2009-03-03","publish_status":"0","recid":"3004","relation_version_is_last":true,"title":["トリパノソーマ病特異的診断法の開発および野外材料を用いた実用評価"],"weko_creator_id":"12","weko_shared_id":-1},"updated":"2024-05-20T01:13:15.410445+00:00"}