3-磷酸甘油醛脱氢酶(GAPD)动力学方法的初步研究  

Preliminary study of a kinetic method of Glyceraldehyde-3-phosphate dehydrogenase determination

作　　者：时娟[1] 张一兵[2] 罗喜钢[2] 王毅[2] 赵园园[1] 

机构地区：[1]辽宁医学院,辽宁锦州121001 [2]辽宁医学院附属第三医院,121000

出　　处：《中国医学工程》2011年第10期14-16,共3页China Medical Engineering

摘　　要：目的建立一种简便、快捷、准确的GAPD的检测方法,为以后的临床推广奠定基础。方法以三乙醇胺作为缓冲液(PH为7.4),1,3-二磷酸甘油酸为底物,偶联3-磷酸甘油酸激酶,还原型辅酶Ⅰ(NADH)被氧化成氧化型辅酶Ⅰ(NAD+),在波长340nm处检测吸光度的变化值,在一定范围内,吸光度的下降与GAPD的活性成正比,间接地计算出GAPD的活性。结果本测定方法的最适PH为7.4,米氏常数(Km)值为6.3×10-4mmol/L,最适基质浓度为0.007mmol/L,批内、批间变异系数为1.60%、1.94%,回收率为96.6%。在0-140U/L范围内线性关系良好,参考值的范围是4-28U/L(逆向法)。结论本法能够准确快速的测定GAPD的活性,是用各种生化自动分析仪。【Objective】 To establish a simple rapid and accurate kinetic monitoring assay for detecting Glyceraldehyde-3-phosphate dehydrogenase（GAPD）.【Methods】 1,3-diphosphoglycerate was as substrate,coupling 3-phosphoglyceric phosphokinase,GAPD catalyzed the substrate in triethanolamine buffe,after a series of enzyme pair association response;NADH＋H＋ was oxidized to NAD＋.The activity of GAPD was determined at 340nm.【Results】 The most appropriate PH is 7.4 and Km is 6.3×10-4mmol/L;the optional substrate concentration was 0.007mmol/L.Inter-assay coefficient of variation（CV） value was 1.60% and intra-assay CV was 1.94%.The linearity reached 0-140U/L.The normal reference value（180 healthy subjects） was 4-28U/L.【Conclusion】 This method for GAPD determination is rapid,accurate and suitable for any kind of automatic biochemistry analyze.

关 键 词：3-磷酸甘油醛脱氢酶 动力学方法 自动化分析 

分 类 号：R963[医药卫生—微生物与生化药学]