滇金丝猴粪便类固醇激素抽提及检测方法评估  

Evaluation on Extraction and Assay Methods for Fecal Steroid Hormones in Yunnan Snub-Nosed Monkeys

作　　者：黎嘉欣 杨秋立 普天春[2] 卢雁平[2] 王伟[2] 刘学锋[2] 刘定震[1] Li Jiaxin;Yang Qiuli;Pu Tianchun;Lu Yanping;Wang Wei;Liu Xuefeng;Liu Dingzhen(College of Life Sciences,Key Laboratory for Biodiversity Sciences andEcological Engineering of Ministry of Education, Beijing Normal University,Beijing,100875,China;Beijing Key Laboratory of Captive Wildlife Technologies,Beijing Zoo,Beijing,100044,China)

机构地区：[1]北京师范大学生命科学学院,教育部生物多样性与生态工程重点实验室,北京100875 [2]圈养野生动物技术北京市重点实验室,北京动物园,北京100044

出　　处：《野生动物学报》2018年第3期507-512,共6页CHINESE JOURNAL OF WILDLIFE

基　　金：北京市公园管理中心项目《三种金丝猴异地保护的繁育技术研究》(No.ZX2016015);北京师范大学本科生创新基金项目(2015);科技部重点研发计划项目(2016YFC0503200)共同资助。

摘　　要：采用非损伤性取样法监测野生动物的生理状态已被广泛接受,但分析结果却因对粪便样品的抽提和检测方法不同而异。本文以圈养滇金丝猴为研究对象,分别采用乙醇加热法、乙醇-蒸馏水法和乙醇-丙酮法对其粪便样品进行激素抽提,然后采用放射免疫分析法(RIA)和酶联免疫吸附测定法(ELISA)检测其雌二醇(E_2)和孕酮(P_4)水平。结果表明,不论采用RIA还是ELISA检测,选用乙醇加热法和乙醇蒸馏水法所抽提样品的孕酮和雌二醇测定结果均不存在显著差异,而采用乙醇丙酮法所抽提样品的孕酮测定结果显著低于前面两种方法所测定的结果。综合考虑平均含量、相对误差、变异系数及抽提步骤的可行性,建议选用乙醇加热法抽提和RIA检测滇金丝猴粪样中孕酮与雌二醇水平。Non-invasive sampling has been commonly used in monitoring the endocrine status of wild animals in recent years,yet the results vary greatly with different usage of hormone extracting and assay methods.We collected feces of three captive adult female Yunnan snub-nosed monkeys(Rhinopithecus bieti)at Beijing Zoo,and examined and compared the fecal concentrations of estradiol(E 2)and progesterone(P 4)of 21 fecal samples by using three different steroid hormone extracting methods,i.e.,ethanol-heating(EH),ethanol-distilled-water(EW),and ethanol-acetone methods(EA),and two assay methods of radioimmunoassay(RIA)and enzyme-linked immunosorbent assay(ELISA),respectively.We found no significant difference in the fecal concentrations in E 2 and P 4 levels by either EH or EW regardless of using RIA or ELISA,yet P 4 values were lower when estimated by EA than when measured by either RIA or ELISA.In considering the average concentration,relative error,coefficient variation as well as practicability,we recommend to use the ethanol-heating method(EH)to extract and RIA for examining the fecal concentrations of E 2 and P 4 in Yunnan snub-nosed monkeys.

关 键 词：滇金丝猴 类固醇激素 抽提方法 酶联免疫吸附法 放射免疫分析法 

分 类 号：Q57[生物学—生物化学]