酶免法检测普氏野马粪便性类固醇激素的最佳稀释比例  

Optimal Dilution Ratio of Detecting Fecal Steroids of Przewalski's Horse by ELISA Assay

作　　者：葛兴芳[1] 胡德夫[1] 吉晟男[1] 刘刚[1] 张文博[1] 王伯君[1] 曹杰 

机构地区：[1]北京林业大学生物科学与技术学院濒危物种非损伤研究技术实验室,北京100083 [2]新疆普氏野马繁育中心,新疆乌鲁木齐830000

出　　处：《干旱区研究》2016年第1期193-196,共4页Arid Zone Research

基　　金：野生动物保护管理项目"野马放归自然区域生境维护与改造试点"(林规发2013)

摘　　要：甲醇对粪便性类固醇激素的检测有一定的影响,为了确定应用酶联免疫方法（酶免法）检测普氏野马粪便性激素的最佳稀释比例,对甲醇保存的提取样品在检测前用PBS缓冲液进行一定比例的稀释。结果显示：检测孕酮（P）,样本稀释15倍时;检测睾酮（T）,样本稀释15倍时;检测雌二醇（E2）,样本稀释30～50倍时,酶标抗原的结合率（B/B0）接近50%,意味着50%的包被抗体被酶标抗原结合,此时的检测结果较灵敏。Przewalski＇s horse is the world＇s endangered species. There are less than 2 000 in the world,and the vast majority of them are captive or semi-wild individuals. It is of great significance to study the breeding and development of Przewalski＇s horse for the rejuvenation and returning to wild state of this species. Methanol has particular effect on detecting fecal steroids. To determine optimal dilution ratio when using ELISA method to detect fecal steroids of Przewalski＇s horse,we used PBS buffer to dilute the extracted samples which stored in methanol at a series ratios before detecting. The results showed when detecting progesterone（ P）,sample was diluted 15 times; detecting testosterone（ T）,sample was diluted 15 times; detecting estradiol（ E2）,sample was diluted 30-50 times,the enzyme-labelled antigen binding rate B / B0（ %） closed to 50%. It means that 50% coating antibody was bound by the enzyme-labelled antigen,and at that time detecting results were more sensitive. The result provides support in the way for researching breeding and development of non-invasive species.

关 键 词：普氏野马 粪便性激素 甲醇 酶联免疫 稀释比例 

分 类 号：S863[农业科学—野生动物驯养] Q503[农业科学—畜牧兽医]