| 家蚕微孢子虫抗体免疫荧光检测方法的建立及应用 |
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| 引用本文： | 李艳红,谢俪,潘国庆,吴正理,庞敏,周泽扬.家蚕微孢子虫抗体免疫荧光检测方法的建立及应用[J].西南农业大学学报,2006,28(6):990-993. |
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| 作者姓名： | 李艳红 谢俪 潘国庆 吴正理 庞敏 周泽扬 |
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| 作者单位： | [1]西南大学蚕桑重点实验室,重庆400716 [2]重庆师范大学动物学实验室,重庆400047 |
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| 基金项目： | 国家自然科学基金资助项目（30400328,30371085,30600452）;科技部“973”重点资助项目（2005CB121000）;国家茧丝办风险基金资助项目（M012005-000 Y-00070） |
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| 摘 要： | 通过制备家蚕微孢子虫总蛋白多克隆抗体，建立了多克隆抗体免疲荧光检测方法（IFA），并利用此方法特异性地检测出了体外培养细胞系（家蚤胚胎细胞系BmE—SWUI）中处于不同发育阶段的家蚤微孢子虫。结果在细胞爬片中可观察到呈较强的黄绿色荧光微孢子虫以及在BmE—SWU1细胞中的寄生分布情况，还可看到孢子极丝的弹出等现象。IFA检测方法鉴别诊断家蚕微粒子虫具有快速、特异性强、灵敏度高、稳定性好、操作简便等特点，可以广泛应用于微孢子虫形态学、流行病学以及蛋白质的定位等研究领域。
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| 关 键 词： | 家蚕微孢子虫 免疫荧光技术 IFA 多克隆抗体 |
| 文章编号： | 1000-2642（2006）06-0990-04 |
| 收稿时间： | 09 4 2006 12:00AM |
| 修稿时间： | 2006-09-04 |
ESTABLISHMENT AND APPLICATION OF INDIRECT IMMUNOFLUORESCENCE ASSAY USING POLYCLONAL ANTIBODY AGAINST NOSEMA BOMBYCIS |
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| LI yan-hong,XIE Li,PAN Guo-qing,WU Zheng-li,PANG Min,ZHOU Ze-yang.ESTABLISHMENT AND APPLICATION OF INDIRECT IMMUNOFLUORESCENCE ASSAY USING POLYCLONAL ANTIBODY AGAINST NOSEMA BOMBYCIS[J].Journal of Southwest Agricultural University,2006,28(6):990-993. |
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| Authors: | LI yan-hong XIE Li PAN Guo-qing WU Zheng-li PANG Min ZHOU Ze-yang |
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| Institution: | 1. The Key lahoratory of Sericuhure of Agrieuhural ministry, Southwest University, Chongqing 400712, China; 2. College of Bioseienee of Chongqing Normal University, Chnngqing 400047, China |
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| Abstract: | An indirect fluorescent-antibody assay(IFA) was established to detect Nosema bombycis with polyclonal antibody prepared against total proteins extracted from germinated N.bombycis.By IFA,antisera against N.bombycis reacted strongly with the mature spores collected from silkworm and with the spores in the cell culture(BmE dell-line),and obvious distinction was observed between the spores and the host cell for their intensity of fluorescent light.The polar tube of N.bombycis was also noticed.These results demonstrated that polyclonal antisera of N.bombycis can be used for indirect immunofluorescence antibody staining to detect N.bombycis and that IFA is a simple,stable and fast method with good specificity.Therefore,IFA can be used in the research fields of the morphology,epidemiology and protein localization of N.bombycis. |
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| Keywords: | Nosema bombycis immunofluorescence IFA polyelonal antibody |
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