Development of rapid immunochromatographic assay as a diagnostic tool for bovine cryptosporidiosis
| Project/Area Number |
19F19107
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| Research Category |
Grant-in-Aid for JSPS Fellows
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| Allocation Type | Single-year Grants |
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| Section | 外国 |
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| Review Section |
Basic Section 42020:Veterinary medical science-related
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| Research Institution | Obihiro University of Agriculture and Veterinary Medicine |
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Principal Investigator |
西川 義文 帯広畜産大学, 原虫病研究センター, 教授 (90431395)
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| Co-Investigator(Kenkyū-buntansha) |
FEREIG RAGAB 帯広畜産大学, 原虫病研究センター, 外国人特別研究員
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| Project Period (FY) |
2019-04-25 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2020: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2019: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | クリプトスポリジム / ウシ / 下痢症 / 診断 |
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| Outline of Research at the Start |
本研究の目的は、公衆衛生および家畜衛生上の重大な問題となっているクリプトスポリジウムに対する新規診断システムを構築し、疫学調査を通じて社会実装につなげることにある。本研究ではプロテオーム解析により優れた診断用抗原を新たに同定し、それら抗原を搭載したイムノクロマトテストの確立を目指す。本診断技術は急性感染期の抗原検出、慢性感染期の抗原虫抗体の検出を想定しており、両検査システムを疫学調査に導入することでエンデミックとエピデミック感染を把握することが可能となる。エジプトでのクリプトスポリジム感染の関心は極めて高く、本研究成果により日本・エジプトの本格的な二国間共同研究につながることが期待される。
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| Outline of Annual Research Achievements |
Hybridoma cells were prepared by fusion of SP2 myeloma cells with the spleen of mice showing high production of antibody as confirmed by ELISA and indirect immunofluorescence test (IFAT). Hybridoma cells were screened by IFAT and ELISA to detect antibody production. Hybridoma cells producing potent and specific antibody were used for immunization of SCID mice for expansion of antibody production in ascites. Further testing of monoclonal antibody (mAb) in ascites was applied using IFAT, ELISA and western blotting. 3 clones out from 360 showed high proficiency and specificity in detection of C. parvum oocysts, lysate antigen and/or rCpGP15. However, our main goal will be focused on evaluation of these mAbs using immunochromatographic test (ICT) to detect C. parvum antigens in cattle feces. Validation of our developed ICTs was applied against fecal samples of cattle previously tested with commercial ICT. Only 1 monoclonal antibody namely D1-8 that prepared from immunization of mice with viable oocysts showed a moderate efficacy for detection of fecal samples from naturally infected calves as confirmed by commercial ICT and light and fluorescent microscopy. The band intensity of ICT was correlated positively with high concentration of used oocyst lysate, with no reactivity to PBS, RIPA buffers. Due to economic and public health importance, the successful development of ICT for detection of C. parvum in calf feces will be an essential addition to Cryptosporidium research because of providing novel knowledge concerning the methodology and preparation of ICT technology.
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| Research Progress Status |
令和2年度が最終年度であるため、記入しない。
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| Strategy for Future Research Activity |
令和2年度が最終年度であるため、記入しない。
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Report
(2 results)
Research Products
(11 results)
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[Journal Article] Toxoplasma gondii Dense Granule Proteins 7, 14, and 15 Are Involved in Modification and Control of the Immune Response Mediated via NF-κB Pathway.2020
Author(s)
Ihara F, Fereig RM, Himori Y, Kameyama K, Umeda K, Tanaka S, Ikeda R, Yamamoto M, Nishikawa Y.
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Journal Title
Front Immunol.
Volume: 11
Pages: 1709-1709
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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