Establishment of a reference ELISA for measurement of universal thresholds of pregnanediol glucuronide excretion rates using urine samples diluted to a constant volume per unit time.
Establishment of a reference ELISA for measurement of universal thresholds of pregnanediol glucuronide excretion rates using urine samples diluted to a constant volume per unit time.
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DOI：
10.1080/15321819.2016.1236730
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Blackwell, Leonard F
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作者：
Binnie, Jan E;Cooke, Delwyn G;Blackwell, Leonard F
描述了一种用于测量分析前稀释至150 mL/h的尿液样本中孕甾烷-3 α-葡糖苷酸（PdG）排泄率的酶联免疫吸附试验（ELISA）。9条优化的标准曲线的灵敏度为0.093 ± 0.070 mumolPdG/24 hr，多重组合标准曲线的平均中点（EC_（50））为6.88 mumolPdG/24 hr，PdG阈值排泄率为7.0 mumol/24 hr，可作为生育结束的标志，位于标准曲线的最准确区域。ELISA的特异性使用正态变量转换来确定，以比较用ELISA方法获得的7个月经周期曲线与先前使用经验证的放射免疫测定（RIA）方法获得的曲线。循环曲线均在实验误差范围内一致，并且使用德明回归获得了高度相关性。相关方程为Y = 1.57X-0.11 μ molPdG/24小时（n = 200; r = 0.932）。ELISA法测定的PdG排泄率比RIA法高50%，但正常范围与原参考气液色谱法相似。因此，ELISA测定法适合作为测量PdG排泄率阈值的参考方法。
An enzyme-linked immunosorbent assay (ELISA) for measurement of pregnanediol-3alpha-glucuronide (PdG) excretion rates in urine samples diluted to 150 mL/h before analysis is described. The sensitivity of the 9 optimized standard curves was 0.093 ± 0.070 mumol PdG/24 hr, with the multiple combined standard curves having a mean mid-point (EC50) of 6.88 mumol PdG/24 hr. The PdG threshold excretion rate of 7.0 mumol/24 hr, which is used as a marker for the end of fertility, was situated in the most accurate region of the standard curve. The specificity of the ELISA was determined using normal variate transformation to compare seven menstrual cycle profiles obtained with the ELISA method with the profiles obtained previously using a validated radioimmunoassay (RIA) method. The cycle profiles all agreed within experimental error, and a high degree of correlation using Deming regression was obtained. The correlation equation was Y = 1.57X-0.11 mumol PdG/24 hr (n = 200; r = 0.932). The PdG excretion rates determined by the ELISA were 50% higher than given by RIA, but the normal ranges were similar to those given by the original reference gas liquid chromatographic method. The ELISA assay was therefore suitable as a reference method for measurement of thresholds of PdG excretion rates.