【摘要】 目的探讨一种血清铜蓝蛋白亚铁氧化酶活性的新的测定方法,并使实验方法和步骤可用于自动化分析。方法用亚铁离子作底物,37℃pH5.80.45mol/L醋酸盐缓冲液条件下Fe2+被血清铜蓝蛋白亚铁氧化酶氧化成Fe3+,一定时间后,加入亚铁嗪显色,λ570nm下测定未被氧化的Fe2+离子的含量,依据反应前后底物浓度的变化,用逆向两点校正计算出被氧化的Fe2+所对应的亚铁氧化酶活性。结果亚铁嗪与Fe2+显色稳定,效果好。标本平均回收率105.2%,线性可达780U/L;精密度:批内CV:4.8%,批间:5.5%,标本存放4℃下稳定时间可达1周以上;脂血、溶血标本不影响亚铁氧化酶测定;EDTA能完全络合Fe2+离子。测定正常人标本30人份,平均值262.1U/L。SD:69.2U/L。结论运用逆向两点校正法,用EDTA作第二校准物取代铜蓝蛋白标准,克服了酶易失活的弱点,使校准更加可靠。可用于自动化分析,检测速度快、精密度较高,易于推广。更多还原

【Abstract】 Objective To investigate a method of detecting serum caruloplasmin ferroxidase activity for automated analyzer.Methods Using Fe 2+ ions as the substrate,optimun assay recation conditions was installed at0.45mol/L pH5.8acetate buffer.During the catalytic oxidation of ceruloplasmin,Fe 2+ was oxidized to Fe 3+ .The remain-ing nonoxidized ferrous ions were detected photometrically at570nm after the chro-mogen being added.Feroxidase activity was estimated by the difference in the ferrous ion concentration before and after the enzymatic reaction.Results The ferrozine formed a highly colored Fe 2+ complex with ferrous ions.The mean recovery was105.2%.The upper limit of linearity in the assay was780U/L.The precision(CV)of interassay and interassay were4.8%and5.5%respectively.The mean value of serum ceruloplasmin ferroxidase activity in healthy persons was262.1U/L,SD:69.2U/L.Conclu sion EDTA which instead of ceruloplasmin as a calibrator was the second main point in the method of two-point inverse calibration had more advantages.It is a rapid,specific and simple method for detecting the serum ferroxidase activity,and can also be applied by automated analyzer.更多还原