酶偶联比色法测定血清门冬氨酸氨基转移酶

Determinaton of asparate aminotransferase in serum with colorimetric method of enzymatic couple

目的 建立一种酶偶联比色法测定血清门冬氨酸氨基转移酶（ＡＳＴ）的方法。方法 谷氨酸在ＮＡＤ＋存在下，通过谷氨酸脱氢酶脱氢反应生成ａ－酮戊二酸和ＮＡＤＨ，ＮＡＤＨ在心肌黄酶（ＦＡＤ）的作用下，与硝基四氮唑兰（ＮＢＴ）发生氧化还原反应，使ＮＢＴ生成甲 ，其颜色的深浅与血清门冬氨酸氨基转移酶活性成正比。结果 该方法的批内精密度为（ＣＶ）３．６％，批间精密度（ＣＶ）为 ４．９％，与酶速率法相关良好，ｒ＝ ０．９９８，线性范围 ０～８５０Ｕ／Ｌ。结论 酶偶联比色法是一种准确简便快速的测定血清ＡＳＴ的方法，适合临床应用。

Objective To develop a colorimetric method of enzymatic couple for asparate aminotrasferase. Methods Glutamate was converted to the a - Ketoglutaric acid and NADH in the presense of NAD+ by glutamic dehydrogense. The NADH formed reacts with nitrotetrazolium blue( NBT) in a diaphorase catalyzed reaction to form a formazan dye, The colour in tensity of which is propotional to the AST activity in the sample. Results This method has a remarkably good correlation with theenzymatic coupling method( r = 0.998); the within - run and between - run( CV%) are less than 3.6 and 4.9 respectively. The linear range of assays is 0～ 850U/L.Conclusion The procedure is simple, rapid and accurate for clinical applies- tion.