Direct enzyme immunoassay of fecal estrone derivatives in dairy cows
Direct enzyme immunoassay of fecal estrone derivatives in dairy cows
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DOI：
10.1111/j.1740-0929.2005.00257.x
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发表时间：
2005-06-01
影响因子：
2
通讯作者：
Nakao, Toshihiko
中科院分区：
文献类型：
--
作者：
Isobe, Naoki;Nakao, Toshihiko
本研究旨在建立一种新的、实用的、简便的检测奶牛粪便中雌酮衍生物(雌酮、雌酮硫酸酯和雌酮葡萄糖醛酸苷)的方法。将粪便与硼酸盐缓冲液混合制成粪液，不经提取直接放入井中，与抗雌酮抗体和辣根过氧化物酶标记雌酮孵育。以硫酸雌酮为标准。该方法的灵敏度为0.15 ng/m L(0.6 ng/g)。批内和批间变异系数分别为5.3-8.1%和13.4-15.7%。回收率为78-102%。只需要4h就可以完成一项测定粪便雌酮衍生物浓度的实验。血浆硫酸雌酮浓度与粪便硫酸雌酮当量浓度呈显著正相关。当测量怀孕奶牛粪便中硫酸雌酮的当量浓度时，从怀孕150天开始逐渐增加，随后从240天到产犊日急剧增加。这些结果表明，本研究建立的直接酶免疫法是一种实用、可靠的检测粪便雌酮衍生物浓度的方法。
The present study was undertaken to develop a novel, practical and simple procedure for enzyme-linked immunosorbent assay of fecal estrone derivatives (estrone, estrone sulfate and estrone glucuronide) in dairy cows. Fecal solution, prepared by mixing feces with borate buffer, was applied directly to wells without extraction, and incubated with anti-estrone antibody and horseradish peroxidase-labeled estrone. Estrone sulfate was used as the standard. The sensitivity of the assay was estimated as 0.15 ng/mL (0.6 ng/g). The intra-assay and inter-assay coefficients of variation were 5.3-8.1% and 13.4-15.7%, respectively. The recovery rate was 78-102%. Only 4 h were needed to complete an assay to measure fecal estrone derivative concentrations. A significant positive correlation was established between plasma estrone sulfate concentrations and fecal estrone sulfate equivalent concentrations. When fecal estrone sulfate equivalent concentrations were measured in pregnant dairy cows, a gradual increase from day 150 of pregnancy, and subsequent drastic increases from day 240 to calving date were observed. These results suggest that the direct enzyme immunoassay procedure developed in the present study is a practical and reliable method for measuring fecal estrone derivative concentrations.