血清葡萄糖及1,5-脱水葡萄糖醇双项同测的建立

Establishement of a dual test method of glucose and 1,5-anhydroglucitol

目的建立新全酶终点法同时测定血清中1，5-脱水葡萄糖醇（1，5-AG）和葡萄糖（Glu）。方法用葡萄糖激酶（GK）和葡萄糖-6-磷酸脱氢酶（G6PD）偶联系统使血清中Glu彻底转化为不与呋喃糖氧化酶（PROD）反应的6-磷酸葡萄糖酸内酯（6-PGA），以其吸光度变化计算出Glu浓度；以PROD氧化1，5-AG，生成1，5-脱水果糖和H2O2，用Trinders反应比色测定出1，5-AG的浓度。结果1，5-AG在550μmol／L（Y=0．001X＋0．0658，r=0．999）以内，Glu浓度在40mmol／L（Y=0．051X＋0．128，r=0．9989）以内线性良好。Glu和1，5-AG的平均批内变异系数（CV）分别为0．88％和1．05％，批间CV分别为1．40％和1．94％。Glu和1，5-AG的平均回收率分别为100．2％和101．6％。三酰甘油（TG）≤8．9mmol／L、血红蛋白（Hb）≤4g／L、总胆红素≤350μmol／L时不影响Glu和1，5-AG的测定。本法测定l，5-AG与Lana微柱法及GK法与HK法测定Glu相关性均良好（r^2=0．999）。Glu与1，5-AG的95％参考值范围分别为3．7—5．7mmol／L和83．1～240．7μmol／L。结论本法简便快速、灵敏、准确和稳定，可自动化双项同测Glu和1，5-AG，为糖尿病的检测与治疗监控提供了新的方法。

Objective To establish a novel enzymatic method for serum glucose and 1, 5-anhydroglueitol （ 1,5- AG） detemination. Methods Linking system of glucokinase （GK） and Glueose-6-phosphate dehydrogenase （ G6PD ） were used to get rid of interference of serum glucose, then calculated glucose concentration according to absorbency variety ,and further,pyranose oxidase （PROD） was used to oxidize 1,5-AG with the formation of 1,5-anhydro fructose and H2 02. The level of 1,5-AG was determined by Trinders reaction principle. Results The linear ranges of glucose and 1,5- AG levels within 40mmol/L （ Y1.5-AG = 0. 001X ＋ 0. 0658, r = 0.999 ） and 550μmoL/L （ YGlu = 0. 051X ＋ 0. 128, r = 0. 998 9） were good. The within-run mean coefficient of variation（CV） of glucose and 1,5-AG were 0.88% and 1.05% ,and between-run were 1.40% and 1.94% , respectively. The average recovery of glucose and 1,5-AG were 100.2% and 101.6%. Triglyeride ≤8.9 mmol/L, hemoglobin≤4 g/L,total bilirubin≤350 μmol/L did not interfere measuring glucose and 1,5-AG. The method has significant correlation with Lana tiny-cotumnination, GK method and HK method. The 95% reference range of glucose and 1,5-AG were 3.7-5.7 mmol/L and 83.1-240.7 μmol/L. Conclusions This dual test method is easy. rapid and precise to diagnose and monitor diabetes mellitus.