奶牛孕酮酶联免疫检测试剂盒的研制  

Preparation of enzyme linked immunosorbent assay kit for milk cow progesterone

作　　者：庞观龙[1] 何达崇[2] 温和心[2] 

机构地区：[1]广西农业职业技术学院,南宁530007 [2]广西科学院生物研究所,南宁530003

出　　处：《广西农业科学》2010年第4期386-389,共4页Guangxi Agricultural Sciences

基　　金：广西自然科学基金项目(桂科基0575028)

摘　　要：为了掌握母牛孕酮水平变化信息,为发情鉴定、妊娠鉴别和卵巢疾病诊断提供参考依据,根据ELISA基本原理,研制了奶牛孕酮ELISA检测试剂盒。结果表明,采用孕酮-牛血清白蛋白(P4-BSA)交联物解决了在酶标板上包被抗原的问题,且抗原容易保存;而利用孕酮-卵清白蛋白(P4-OVA)交联物免疫兔子获得高效价抗体,能阻断1 ng/mL孕酮水平;并以ELISA棋盘测定法确定P4-BSA 2000倍、血清500倍稀释为该试剂盒最佳工作浓度。该试剂盒以牛粪作为检测样品,即通过粪便中的孕酮水平变化可为奶牛发情鉴定、妊娠鉴别、繁殖障碍性疾病诊断提供可靠的技术支撑。In order to investigate the change in progesterone level of cow to provide reference on heat and gestation period identification, and ovary disease diagnosis, the enzyme linked immunosorbent assay kit（ELISA） for cow progesterone was prepared based on primary mechanism of ELISA. The results showed that applying progesterone bovine serum albumin （Pa-BSA） cross-linking polymer could solve the problem of enveloping antigen on ELISA plate, and antigen was easy to be conserved. It was found that high titer antibody could be obtained by applying progesterone-ovalbumin（P4-OVA） cross-linking polymer to immunize the rabit and it could block 1 ng/mL progesterone. The 2000 and 500 dilution times for P4- BSA and serum were found to be the optimal concentration of kit by ELISA chessboard method. The cow dung was used for detecting sample with kit, and the change in progesterone level of dung could provide reference on heat and gestation period identification, and diagnosis of reproduction barrier disease.

关 键 词：奶牛 孕酮 酶联免疫吸附测定法 抗体效价 

分 类 号：S854.43[农业科学—临床兽医学]