{"created":"2023-05-15T15:17:00.391082+00:00","id":3006,"links":{},"metadata":{"_buckets":{"deposit":"814640c3-fade-4bce-bea0-b42cfcc3672d"},"_deposit":{"created_by":12,"id":"3006","owners":[12],"pid":{"revision_id":0,"type":"depid","value":"3006"},"status":"published"},"_oai":{"id":"oai:obihiro.repo.nii.ac.jp:00003006","sets":["242:243"]},"author_link":["58","5361","340","295","63","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 international standard diagnostic method for equine babesiosis","subitem_alternative_title_language":"en"}]},"item_6_contributor_5":{"attribute_name":"研究分担者","attribute_value_mlt":[{"contributorNames":[{"contributorName":"藤崎, 幸蔵"}],"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"}]},{"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":[{"familyName":"Xuan","familyNameLang":"en"},{"familyName":"玄","familyNameLang":"ja"}],"givenNames":[{"givenName":"Xuenan","givenNameLang":"en"},{"givenName":"学南","givenNameLang":"ja"}],"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"}]},{"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":[{"familyName":"Yokoyama","familyNameLang":"en"},{"familyName":"横山","familyNameLang":"ja"}],"givenNames":[{"givenName":"Naoaki","givenNameLang":"en"},{"givenName":"直明","givenNameLang":"ja"}],"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":[{"contributorAffiliationScheme":"ISNI","contributorAffiliationURI":"http://www.isni.org/isni/"}],"contributorAffiliationNames":[{"contributorAffiliationNameLang":"ja"}]}],"contributorNames":[{"contributorName":"Inoue, Noboru","lang":"en"},{"contributorName":"井上, 昇","lang":"ja"}],"familyNames":[{"familyName":"Inoue","familyNameLang":"en"},{"familyName":"井上","familyNameLang":"ja"}],"givenNames":[{"givenName":"Noboru","givenNameLang":"en"},{"givenName":"昇","givenNameLang":"ja"}],"nameIdentifiers":[{"nameIdentifier":"63","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"9000002841346","nameIdentifierScheme":"CiNii ID","nameIdentifierURI":"http://ci.nii.ac.jp/nrid/9000002841346"},{"nameIdentifier":"10271751","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=10271751"},{"nameIdentifier":"read0185948","nameIdentifierScheme":"researchmap","nameIdentifierURI":"https://researchmap.jp/read0185948"}]},{"contributorNames":[{"contributorName":"筏井, 宏実","lang":"ja"}],"nameIdentifiers":[{"nameIdentifier":"5361","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"80327460","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=80327460"}]}]},"item_6_description_10":{"attribute_name":"研究代表者番号","attribute_value_mlt":[{"subitem_description":"80159582","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_type":"Other"}]},"item_6_description_13":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"本研究はウマバベシア病に対する感度と特異性の高い国際標準診断法を開発・実用化することを目標として、Babesia equiとB.caballiの組換え原虫タンパク質の作製、免疫原性の解析、組換え原虫タンパク質を用いたELISA、イムノクラマト法の確立、世界各国の血清試料を用いた血清診断法の評価を行い、下記のような成果を得た。\n1.バベシア原虫の新規の抗原\n感染ウマから得られた血清を用いたクローニングにより、新規のB.equi Be82、Be158遺伝子およびB.caballi Bc134遺伝子が得られた。大腸菌で発現された原虫タンパク質をELISA用抗原として用いたところ、高い特異性および感度が認められた。\n2.ウマバベシア原虫組換え抗原産生の検討\nB.equiのメロゾイト表面抗原(EMA-1)およびB.caballiのロプトリー抗原の組み換抗原(BC48)を大腸菌を用いて産生し、発現された原虫タンパク質は分子量、抗原性など原虫由来のものと一致することが確認された。\n3.組換え抗原を用いたELISA\nB.equiのメロゾイト表面抗原(EMA-2)およびB.caballiのロプトリー抗原の組み換え抗原(BC48)を用いたELISAは感度及び特異性に優れていることが示された。また、野外血清を用いた試験でも、同様の結果が得られた。\n4.イムノクロマト法の検討\n高濃度に発現されたB.equiのEMA-2およびB.caballiのBC48を用いて、特異的反応が高く、短時間で判定可能なイムノクロマト法の開発に成功した。また、これら2種類の抗原を一つのスティックに固定して、同時に2種類のバベシア感染を診断できるキットも開発された。\n以上のように本研究では、B.equiのメロゾイト表面抗原(EMA-2)およびB.caballiのBC48が最も診断用抗原として優れていることが判明した。また、両抗原を用いたELISAおよびイムノクロマト法は特異性および感度に優れていることが示された。今後世界的な比較実験及び評価を行うことによって、国際的標準診断法の診断用抗原として使用される可能性が極めて高いと期待される。","subitem_description_language":"ja","subitem_description_type":"Abstract"},{"subitem_description":"Equine babesiosis, caused by Babesia equi and Babesia caballi, is characterized by fever, anemia, icterus, hepatosplenomegaly, lethargy, and, in some cases, leading to great economic losses in the horse industry. The infected horses often remain carriers of the parasites for a long period and are known to act as sources for subsequent infections to other horses via tick vectors. Therefore, the development of a high-quality system has been required for the serological diagnosis of babesial infection. In the present study, development of international standard diagnostic methods with high specificity and sensitivity for equine babesiosis was examined by gene cloning of Babesia DNA with immune sera, production of recombinant antigens, evaluation of ELISA and imunochromatographic test using those antigens. Novel genes, Be82, Be158 and Bc134, were cloned from B.equi and B.caballi, respectively. ELISA using these recombinant antigens showed the high specificity and sensitivity for the detection of antibodies against both babesial infections. Productions of recombinant antigens in Escherichia coli, EMA-2 and BC48, were also improved by the construction of truncated genes or modification of culture or induction factors. ELISA using these truncated EMA-2 and BC48 showed high specificity and sensitivity for the detection of antibodies. An immunochromotographic test for simultaneous detection of both B.caballi and B.equi specific antibodies was developed. Evaluation in the detection of sera from uninfected horses and experimentally infected horses showed high sensitivity and specificity as shown in ELISA. A differential single-round and multiplex polymerase chain reaction (PCR) method was also developed for the simultaneous detection of B.caballi and B.equi, by targeting 18S ribosomal RNA genes. In conclusion, EMA-2 for B.equi and BC48 for B.caballi were most excellent as antigens for the diagnosis. ELISA and immunochromatographic test using both antigens have shown to have high specificity and sensitivity for the detection of antibodies. The worldwide experimental comparison and evaluation of these serological tests and genetic analysis will would be feasible tests for internationally recognized diagnosis for equine babesiosis in the world.","subitem_description_language":"en","subitem_description_type":"Abstract"}]},"item_6_description_14":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"2001年度～2004年度科学研究費補助金基盤研究(A)(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":"13356007","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":"I090303-6.pdf","filesize":[{"value":"19.5 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"I090303-6.pdf","url":"https://obihiro.repo.nii.ac.jp/record/3006/files/I090303-6.pdf"},"version_id":"c501b94d-4a23-4564-bff9-00df650a53f0"}]},"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":[{"affiliationNameIdentifier":"","affiliationNameIdentifierScheme":"ISNI","affiliationNameIdentifierURI":"http://www.isni.org/isni/"}],"affiliationNames":[{"affiliationName":"","affiliationNameLang":"ja"}]}],"creatorNames":[{"creatorName":"Igarashi, Ikuo","creatorNameLang":"en"},{"creatorName":"五十嵐, 郁男","creatorNameLang":"ja"}],"familyNames":[{"familyName":"Igarashi","familyNameLang":"en"},{"familyName":"五十嵐","familyNameLang":"ja"}],"givenNames":[{"givenName":"Ikuo","givenNameLang":"en"},{"givenName":"郁男","givenNameLang":"ja"}],"nameIdentifiers":[{"nameIdentifier":"58","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"9000003078567","nameIdentifierScheme":"CiNii ID","nameIdentifierURI":"http://ci.nii.ac.jp/nrid/9000003078567"},{"nameIdentifier":"80159582","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=80159582"},{"nameIdentifier":"read0167555","nameIdentifierScheme":"researchmap","nameIdentifierURI":"https://researchmap.jp/read0167555"}]}]},"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":"ウマバベシア病に対する国際標準診断法の開発","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"ウマバベシア病に対する国際標準診断法の開発","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":"3006","relation_version_is_last":true,"title":["ウマバベシア病に対する国際標準診断法の開発"],"weko_creator_id":"12","weko_shared_id":-1},"updated":"2023-12-07T04:32:09.983933+00:00"}