Evaluation of a kinetic uricase method for serum uric acid assay by predicting background absorbance of uricase reaction solution with an integrated method.
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.
Liao
Fei
F
Laboratory of Bioinformatics and Molecular Engineering, Chongqing University of Medical Sciences, Chongqing 400016, China. liaofeish@vip.sina.com
Zhao
Yun-sheng
YS
Zhao
Li-na
LN
Tao
Jia
J
Zhu
Xiao-yun
XY
Liu
Lan
L
D006801:Humans; D007700:Kinetics; D011933:Reagent Kits, Diagnostic; D013056:Spectrophotometry, Ultraviolet; D014503:Urate Oxidase / Q000737:chemistry*; D014527:Uric Acid / Q000097:blood* / Q000378:metabolism
D011933:Reagent Kits, Diagnostic; D014527:Uric Acid; D014503:Urate Oxidase