以WST-5为色源脱氢酶法测定血清1,5-AG含量  

A new enzymaitic method for determination of 1,5-anhydro-D-glucitol in serum using WST-5 as a color source

作　　者：潘利琴[1] 张德亭[1] 连国军[2] 夏淑琦[1] 曹建明[2] 马美萍[2] 

机构地区：[1]浙江省温州市第三人民医院检验科,浙江温州325000 [2]温州医学院环境与公共卫生学院,浙江温州325000

出　　处：《中国卫生检验杂志》2013年第3期688-690,共3页Chinese Journal of Health Laboratory Technology

基　　金：浙江省科技厅公益技术研究社会发展项目(2011C33043);温州市科技计划项目(Y20090247)

摘　　要：目的:建立以WST-5为色源,脱氢酶法测定血清1,5-AG含量的新方法。方法:用ATP依赖性己糖激酶将1,5-脱水葡萄糖醇和主要干扰物葡萄糖分别转化为1,5-脱水葡萄糖醇-6-磷酸和葡萄糖-6磷酸,转化后的1,5-脱水葡萄糖醇-6-磷酸被1,5-脱水葡萄糖醇-6-磷酸脱氢酶脱氢氧化,同时使水溶性WST-5还原为有色物质,比色定量。结果:该法线性范围为0μmol/L～504μmol/L,平均回收率为99.3%;批内、批间相对标准差(RSD)分别为1.2%～3.3%和2.4%～3.9%;与参考方法相比的回归方程及相关系数分别为Y=1.009X-1.117,r=0.9984。结论:该法具有快速、简便、灵敏可靠等优点,适合临床应用。Objective：To establish a new enzymaitic method for determination of 1,5 - anhydro - D - glucitol in serum using WST-5 as a color source. Methods： The 1,5 -anhydro- D -glucitol （1,5 -AG） which was object to be measured and the glucose （Glu） which was the main interference were converted to 1,5 -dehydrated glucose - 6 - phosphoric acid （ 1,5 - AG - 6 - P） and glucose - 6 phosphoric acid （ Glu - 6 - P） by the presence of ATP - dependent Hexokinase （ ATP - HK）. The 1,5 - AG - 6 - P was oxidized by 1,5 - anhydro - D - glucitol - 6 - phosphate dehydrogenase, with the formation of a color - dye, the absorbence of the color - dye was quantified with the concentration of 1,5 - anhydro - D - glucitol. Results： The method was linear in the range of 0 μmol/L - 504 μmol/L, the average rate of recovery was 99.3% , the within - run and between - run RSDs were 1.2% - 3.3% and 2.4% - 3.9% respectively. Comparing this method （Y） with the reference method （ X）, the regression equation was Y = 1. 009X - 1.117, r = 0. 9984. Conclusion： This method was simple, sensitive and suitable for clinical application.

关 键 词：1 5-脱水葡糖醇 1 5-脱水葡糖醇-6-磷酸脱氢酶 WST-5 

分 类 号：R446.1[医药卫生—诊断学]