Satterlee D G, Abdullah R B, Gildersleeve R P
Poult Sci. 1980 Apr;59(4):900-5. doi: 10.3382/ps.0590900.
The objective was to develop and validate a rapid, precise plasma corticosterone radioimmunoassay using a commercially available antiserum, for use in Gallus domesticus. Sample preparation consisted of sequential 2,2,4-trimethylpentane and dichloromethane extraction to partition progestins and glucocorticoids, respectively. Progesterone, 11 beta-Hydroxyprogesterone, and deoxycorticosterone significantly cross reacted with the antiserum used. However, progesterone was effectively removed prior to assay by the 2,2,4-trimethylpentane plasma washings while 11 beta-Hydroxyprogesterone and deoxycorticosterone interference is doubtful for reasons discussed. Standard curve data showed a linear range from 0 to 150 pg by log-logit transformation with a 5 pg assay sensitivity. Mean percent unlabeled corticosterone recovery was 96% with intraassay and interassay coefficients of variation being 3.75 and 5.62%, respectively. The assay was utilized to characterize corticosterone fluctuations during day one post hatch in broiler chicks. Method of blood collection, rapid decapitation vs. heart stab, resulted in no difference in mean plasma corticosterone levels. Corticosterone levels differed over a 24 hr sampling period, such that highest levels were found upon receipt of the chicks at the hatchery (approximately 20 ng/ml) and 20 hr later (approximately 11 ng/ml). Lowest plasma corticosterone concentrations occurred from 10 to 14 hr (approximately 6 ng/ml) after receipt of the chicks.
目的是开发并验证一种使用市售抗血清的快速、精确的血浆皮质酮放射免疫测定法，用于家鸡。样品制备包括依次用2,2,4-三甲基戊烷和二氯甲烷萃取，分别分离孕激素和糖皮质激素。孕酮、11β-羟基孕酮和脱氧皮质酮与所用抗血清有显著交叉反应。然而，在测定前通过2,2,4-三甲基戊烷血浆洗涤有效地去除了孕酮，而11β-羟基孕酮和脱氧皮质酮的干扰因所讨论的原因存疑。标准曲线数据经对数-对数转换显示线性范围为0至150 pg，测定灵敏度为5 pg。未标记皮质酮的平均回收率为96%，批内和批间变异系数分别为3.75%和5.62%。该测定法用于表征肉鸡雏鸡出壳后第一天皮质酮的波动情况。采血方法，快速断头法与心脏穿刺法，导致平均血浆皮质酮水平无差异。在24小时的采样期内皮质酮水平有所不同，在孵化场收到雏鸡时（约20 ng/ml）和20小时后（约11 ng/ml）发现最高水平。收到雏鸡后10至14小时血浆皮质酮浓度最低（约6 ng/ml）。
