| Project/Area Number |
10839015
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
動物臨床医学
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| Research Institution | Kitasato University |
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Principal Investigator |
KAWAMURA Seiichi. Dep of Veterinary Internal Medicine, Kitasato University, Professor, 獣医畜産学部・獣医内科学講座, 教授 (60050530)
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| Co-Investigator(Kenkyū-buntansha) |
HOSHI. Fumio. Dep of Veterinary Internal Medicine, Kitasato University, Assistant Professor, 獣医畜産学部・獣医内科学講座, 講師 (00219164)
HIGUCH Seiichi Dep of Small Animal Medicine, Kitasato University, Professor, 獣医畜産学部・小動物臨床学講座, 教授 (60095510)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 1999: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1998: ¥3,400,000 (Direct Cost: ¥3,400,000)
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| Keywords | Theileria sergenti / Babesia gibsoni / Diagnosis of protozoa infection / ELISA / Veterinary / Protozoa antigen / Monoclonal antibody to protozoa / Anemia / 原虫感染症 / Babesia ovata / 原虫病の診断液 / 原虫の主要抗原 / 間接蛍光抗体法 |
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| Research Abstract |
1. Preparations of Monoclonal antibodies for protozoan proteins: Mab for protozoa of Babesia spp. And Theileria spp., infected within erythrocytes of cattle and canine were prepared. In a many of Mab for Babesia (B.) gibsoni, there were isolated two Mab recognized proteins of 30 kDa and 24.5 kDa in the protozoa. A Mab for B. ovata recognized proteins of 19 kDa and two Mab for Theileria (T.) sergenti recognized proteins of 23 kDa and 32 kDa.
2. Diagnostic methods using Mab for B. gibsoni
(1) Development of enzyme-linked immunosorbent assay (ELISA) system; ELISA systems by sandwiche method were investigated using each Mab for proteins of 30 and 24.5 kDa in B. gibsoni. In their assay systems, when Mab for 30 kDa protein was used as a antibody of solid phase and Mab for 24.5 kDa protein was labeled with biotin and streptolysin, the sensitivity and specificity of the reaction showed excellent results. There was highly significant correlated between babesial soluble protein in serum of canine
… More
infected with B. gibsoni and percentages of protozoa in erythrocytes (r=0.879). And this ELISA was positive at a percentage above 0.05% parasitemia of protozoa.
(2) Latex agglutination test; Using latex beads sensitized with Mab, the agglutination test using latex sensitized with Mab for 24.5 kDa protein showed most sensitive and specific reaction. The correlation with parasitemia was high significantly (r=0.832). But, this ELISA was negative at a percentage under 0.1% parasitemia of protozoa.
3. Diagnostic methods using Mab for T. sergenti
(1) Development of enzyme-linked immunosorbent assay (ELISA) system; Sandwiche ELISA systems using Mab for 30 and 24.5 kDa in T. sergenti also showed good reactions. In their assay systems, good reactional condition was observed by using Mab for 23 kDa protein.
(2) Latex agglutination test; Using latex beads sensitized with Mab for 23 kDa protein, the agglutination test showed most sensitive and specific reaction. The correlation with parasitemia was high significantly (r=0.801). But, this ELISA was negative at a percentage under 0.2% parasitemia of protozoa. Less
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