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通过一种综合方法预测尿酸酶反应溶液的背景吸光度来评估一种用于血清尿酸测定的动力学尿酸酶法。

Evaluation of a kinetic uricase method for serum uric acid assay by predicting background absorbance of uricase reaction solution with an integrated method.

作者信息

Liao Fei, Zhao Yun-sheng, Zhao Li-na, Tao Jia, Zhu Xiao-yun, Liu Lan

机构信息

Laboratory of Bioinformatics and Molecular Engineering, Chongqing University of Medical Sciences, Chongqing 400016, China.

出版信息

J Zhejiang Univ Sci B. 2006 Jun;7(6):497-502. doi: 10.1631/jzus.2006.B0497.

Abstract

A patented kinetic uricase method was evaluated for serum uric acid assay. Initial absorbance of the reaction mixture before uricase action (A(0)) was obtained by correcting the absorbance at 293 nm measured before the addition of uricase solution, and background absorbance (A(b)) was predicted by an integrated method. Uric acid concentration in reaction solution was calculated from A, the difference between A(0) and A(b), using the absorptivity preset for uric acid. This kinetic uricase method exhibited CV<4.3% and recovery of 100%. Lipids, bilirubin, hemoglobin, ascorbic acid, reduced glutathione and xanthine <0.32 mmol/L in serum had no significant effects. A linearly responded to 1.2 to 37.5 micromol/L uric acid in reaction solution containing 15 microl serum. The slope of linear response was consistent with the absorptivity preset for uric acid while the intercept was consistent with that for serum alone. Uric acid concentrations in clinic sera by different uricase methods positively correlated to each other. By Bland-Altman analysis, this kinetic uricase method accorded with that by quantifying the total change of UV absorbance on the completion of uricase reaction. These results demonstrated that this kinetic uricase method is reliable for serum uric acid assay with enhanced resistance to both xanthine and other common errors, wider range of linear response and much lower cost.

摘要

对一种专利的动力学尿酸酶法进行了血清尿酸测定评估。尿酸酶作用前反应混合物的初始吸光度(A(0))通过校正加入尿酸酶溶液前在293nm处测得的吸光度获得,背景吸光度(A(b))通过积分法预测。反应溶液中尿酸浓度根据A(A(0)与A(b)之差),使用预设的尿酸吸光系数计算得出。这种动力学尿酸酶法的变异系数(CV)<4.3%,回收率为100%。血清中脂质、胆红素、血红蛋白、抗坏血酸、还原型谷胱甘肽和黄嘌呤<0.32mmol/L时无显著影响。在含有15微升血清的反应溶液中,对1.2至37.5微摩尔/升的尿酸呈线性响应。线性响应的斜率与预设的尿酸吸光系数一致,而截距与单独血清的截距一致。不同尿酸酶法测定的临床血清中尿酸浓度彼此呈正相关。通过Bland-Altman分析,这种动力学尿酸酶法与通过量化尿酸酶反应完成时紫外线吸光度的总变化的方法一致。这些结果表明,这种动力学尿酸酶法对于血清尿酸测定是可靠的,对黄嘌呤和其他常见误差的耐受性增强,线性响应范围更广且成本更低。

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