抗贝氏隐孢子虫卵黄抗体的制备与鉴定  

Preparation and identification of yolk antibody against Cryptosporidium baileyi

作　　者：崔朝辉[1] 邵兆霞[1] 菅复春[1] 王荣军[1] 张龙现[1] CUI Zhao-hui SHAO Zhao-xia JIAN Fu-chun WANG Rong-jun ZHANG Long-xian(College of Animal Science and Veterinary Medicine ,Henan Agricultural University/International Joint Laboratory of Zoonosis in Henan Province ,Zhengzhou 45002, Chin)

机构地区：[1]河南农业大学牧医工程学院河南省人兽共患病国际联合实验室,河南郑州450002

出　　处：《中国兽医科学》2017年第7期825-830,共6页Chinese Veterinary Science

基　　金：河南省科技创新杰出人才计划项目(34200510012);十二五国家科技重大专项(2012ZX10004220-011)

摘　　要：为分析抗贝氏隐孢子虫的免疫机制,本研究将纯化的贝氏隐孢子虫卵囊经处理后制备成免疫抗原,经鸡翅下及大腿内侧肌肉多点注射免疫6次150日龄产蛋海兰鸡,每次免疫间隔为15~20 d,以制备特异性抗贝氏隐孢子虫(Cryptosporidium baileyi)卵黄免疫球蛋白(IgY)。结果显示,初次免疫后第120天,IgY效价达到1∶25 600,经免疫荧光法鉴定可识别贝氏隐孢子虫卵囊。用间接ELISA和间接免疫荧光试验比较测定IgY抗体对贝氏隐孢子虫、牛安氏隐孢子虫、猪等孢球虫及猪弓形虫的抗原发现,得到的IgY抗体能特异性识别贝氏隐孢子虫。间接ELISA法测定不同温度、不同pH下IgY的活性发现,制备的IgY抗体具有较好的热稳定性和一定的耐酸、耐碱性。以上结果表明,贝氏隐孢子虫卵囊抗原免疫蛋鸡可制备高质量的特异性IgY抗体,这为进一步应用抗隐孢子虫特异性IgY抗体治疗与诊断隐孢子虫病奠定基础。In order to lay a foundation for analyzing the effect of anti-Cryptosporidium baileyi antibodies,immunoglobulin of yolk（IgY）specifically against Cryptosporidium baileyi was prepared and identified.The purified oocysts of Cryptosporidium baileyi was prepared into immune antigens,and injected into 150-day-old laying hens 6 times under the wings and thigh by muscle multi-point injection,each time interval of immunization was between 15 and 20 d. In results,the IgY titers reached 1 ∶ 25 600 on day 120post-1st-immunization,and C.baileyi oocysts could be specifically recognized by IgY antibody using immunofluorescence.IgY antibody specificity against C.baileyi,Cryptosporidium andersoni, Isospora suis and swine Toxoplasma gondii was comparatively measured by the indirect ELISA and indirect immunofluorescence assay.In result,the obtained IgY antibody could specifically recognize C.baileyi oocysts.Indirect ELISA method was used to determine the activity of different temperature and pH,and the results showed that the obtained antibodies had good thermal stability and acid resistance.Overall,the laying hens could produce IgY antibodies with high quality and specificity after immune by C.baileyi oocysts antigens,which lay a foundation for the further application of anti-Cryptosporidium specific IgY antibodies in diagnosis and treatment of cryptosporidiosis.

关 键 词：贝氏隐孢子虫 特异性IgY抗体 制备 鉴定 

分 类 号：S852.723[农业科学—基础兽医学]