采用夹心 ELISA 法测定 AQP2 的尿排泄日变异性。
Daily variance of urinary excretion of AQP2 determined by sandwich ELISA method.
机构信息
Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan.
出版信息
Clin Exp Nephrol. 2012 Jun;16(3):406-10. doi: 10.1007/s10157-011-0574-2. Epub 2011 Dec 10.
BACKGROUND
Urinary excretion of aquaporin 2 (AQP2) is a useful marker of kidney collecting duct function. A specific and convenient method to measure AQP2 in human urine would help to treat water balance disorders. It is unknown whether urinary excretion of AQP2 shows any daily variance.
METHODS
A sandwich enzyme-linked immunosorbent assay (ELISA) method for AQP2 was established using two different kinds of antibodies, and its sensitivity and specificity were examined. Daily variance of urinary excretion of AQP2 and responses to acute water load were examined.
RESULTS
The established ELISA specifically detected urine AQP2 with high sensitivity (detected as low as 0.34 pmol/mL). Urinary excretion of AQP2 did not show daily variance between 9 a.m. and 9 p.m. in healthy subjects.
CONCLUSIONS
The developed ELISA method using two different antibodies is convenient and highly sensitive, and could be useful in clinical practice. Urinary excretion of AQP2 is relatively constant from morning to night, and spot urine sampling at any time during this time period does not affect the results.
背景
尿中水通道蛋白 2（AQP2）的排泄是反映肾脏集合管功能的有用标志物。一种特异、简便的检测人尿 AQP2 的方法将有助于治疗水液平衡紊乱。但目前尚不清楚 AQP2 的尿排泄是否存在日变异。
方法
采用两种不同的抗体建立了检测 AQP2 的夹心酶联免疫吸附试验（ELISA）方法，并对其敏感性和特异性进行了检测。同时检测了 AQP2 的日排泄变异和对急性水负荷的反应。
结果
该 ELISA 特异性好，检测下限可达 0.34 pmol/ml，对人尿 AQP2 具有较高的敏感性。健康受试者的尿 AQP2 排泄在上午 9 时至晚上 9 时之间无日变异。
结论
本研究建立的使用两种不同抗体的 ELISA 方法简便、敏感，在临床实践中可能具有应用价值。AQP2 的尿排泄从早到晚较为恒定，在此期间的任何时间点采集尿标本均不影响检测结果。
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