Chen S P, Kuan S S, Guilbault G G
Clin Chim Acta. 1980 Jan 1;100(1):21-31. doi: 10.1016/0009-8981(80)90181-3.
An enzymatic semi-solid surface fluorometric method is described for the determination of serum creatinine on silicone-rubber pads. In this method, the glutamate dehydrogenase, alpha-ketoglutarate, ADP and NADH are mixed, then 30 microliters of diluted serum is added. After the free ammonium ion in serum is consumed, creatininase is added to initiate the assay. The rate of disappearance of NADH fluorescence at 460 nm (excitation wavelength 340 nm) is monitored and is proportional to the serum creatinine concentration. The whole assay uses only 0.41 I.U. creatininase, and takes less than 5 min. The calibration curve is linear up to 82 mg creatinine per liter. The proposed method offers a rapid, simple and inexpensive means for creatinine assay. The results obtained correlate well with the modified Jaffe method applied on Technicon SMA 12/60, with a correlation coefficient of 0.998. The recovery averages 99.3%.
描述了一种用于在硅橡胶垫上测定血清肌酐的酶促半固体表面荧光法。在该方法中，将谷氨酸脱氢酶、α-酮戊二酸、ADP和NADH混合，然后加入30微升稀释血清。血清中的游离铵离子被消耗后，加入肌酐酶启动测定。监测460nm（激发波长340nm）处NADH荧光的消失速率，该速率与血清肌酐浓度成正比。整个测定仅使用0.41国际单位的肌酐酶，耗时不到5分钟。校准曲线在每升82毫克肌酐以内呈线性。所提出的方法为肌酐测定提供了一种快速、简单且廉价的手段。所得结果与应用于Technicon SMA 12/60的改良Jaffe法相关性良好，相关系数为0.998。回收率平均为99.3%。
