Development of Diagnostic Systems on the Piroplasma lnfection in Cattle and Canine

牛和犬梨原体感染诊断系统的开发

• 批准号: 10839015
• 负责人: KAWAMURA Seiichi.
• 金额: $ 2.43万
• 依托单位: Kitasato University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10839015/
• 关键词: 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

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

1. 原生动物蛋白单克隆抗体的制备：制备了牛、犬红细胞内感染的巴贝斯虫和希勒氏菌原生动物单克隆抗体。在许多针对巴贝斯虫（B.） gibsoni的Mab单抗中，在原虫中分离到30kda和24.5 kDa的Mab识别蛋白。一个单抗可识别19 kDa蛋白，两个单抗可识别23 kDa和32 kDa蛋白。单抗诊断gibsoni的方法(1)酶联免疫吸附试验（ELISA）系统的建立；采用夹心法，对gibsoni中30和24.5 kDa蛋白的单抗进行ELISA检测。在他们的检测系统中，用30kda蛋白的Mab作为固相抗体，用生物素和溶血素标记24.5 kDa蛋白的Mab，反应的敏感性和特异性都很好。感染gibsoni较多的犬血清中巴贝斯虫可溶性蛋白与红细胞中原虫百分比呈极显著相关（r=0.879）。该酶联免疫吸附试验阳性，原虫寄生率在0.05%以上。(2)乳胶凝集试验；用单抗致敏乳胶珠对24.5 kDa蛋白的凝集试验显示，单抗致敏乳胶珠对24.5 kDa蛋白的凝集反应最为敏感和特异。与寄生虫病的相关性显著（r=0.832）。但在原虫寄生率低于0.1%的情况下，该酶联免疫吸附试验呈阴性。sergenti单克隆抗体诊断方法(1)酶联免疫吸附试验（ELISA）系统的建立；使用30和24.5 kDa单抗的夹心ELISA系统也显示出良好的反应。在他们的检测系统中，用Mab检测23 kDa蛋白，反应条件良好。(2)乳胶凝集试验；用单抗致敏的乳胶珠对23kda蛋白进行凝集试验，反应最灵敏，特异性最强。与寄生虫病的相关性显著（r=0.801）。但该酶联免疫吸附试验在原虫寄生率0.2%以下呈阴性。少

项目成果

Kawamura,S.et al.: "Development of diagnostic methods for Babesiosis and Theileriosis in dogs and cattle." Journal of Veterinary Medical Science. (発表予定).

Kawamura, S. 等人：“狗和牛巴贝斯虫病和泰勒虫病的诊断方法的开发。”《兽医医学科学杂志》（待出版）。

Kawamura,S.et al.: "Monoclonal antibodies for diagnosis of Babesiosis and Theileriosis in dogs and Cattle." Journal of Veterinary Medical Science. (発表予定).

Kawamura, S. 等人：“用于诊断狗和牛巴贝斯虫病和泰勒虫病的单克隆抗体。”《兽医医学科学杂志》（待出版）。