液体型MPT-NAG匀相速率法试剂测定NAG活性  

Determination the NAG activity by new stabile liqulid Homogeneous assay of N-acetyl-beta-D-glucosaminidase with MPT-NAG as Substrate

作　　者：方芳[1] 连国军[1] 赵长容[1] 王文蔚[1] 马美萍[1] 

机构地区：[1]温州医科大学环境与公共卫生学院,浙江温州325000

出　　处：《中国卫生检验杂志》2014年第23期3373-3375,共3页Chinese Journal of Health Laboratory Technology

基　　金：温州市科技计划项目(Y20100194)

摘　　要：目的建立以6-甲基-2-硫代吡啶-N-乙酰-β-D-氨基葡萄糖苷(MPT-NAG)为底物的液体型匀相速率法试剂测定NAG活性。方法在含表面活性剂的柠檬酸缓冲液中,NAG催化MPT-NAG分解产生6-甲基-2巯基吡啶(MPT),用全自动生化分析仪监测340 nm处吸光度的变化速率,与标准液比较后计算出样本中NAG活性。结果该法线性范围为0 U/L^380 U/L,平均回收率为98.8%;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=1.01x+0.24,r=0.9986。结论用液体型MPTNAG匀相速率法试剂测定NAG活性,具有快速、稳定、简便等优点,适合临床应用。Objective To establish a new stabile liqulid homogeneous assay of N-acetyl-beta-D-glucosaminidase with MPT-NAG as Substrate. Methods The analysis is based on the fact that NAG catalytic MPT-NAG decomposition to produce6-methyl-2 mercapto pyridine（ MPT） in citric acid buffer solution containing surfactant,and the rate of absorbance at340 nm is measured by automatic biochemical analyzer. After comparing with standard solution,the result of NAG activity in sample is then calculated. Results The method is liner in the range of 0 U / L - 380 U / L,the average rate of recovery were98. 8%,the within-run and between-run RSDs are 1. 2% - 3. 3 % and 2. 4% - 3. 9 % respectively. Comparing this methods（ y） with the reference method（ x）,the regression equations is y = 1. 01 x ＋ 0. 24,r = 0. 9986. Conclusion This method is rapid,stable,simple and suitable for clinical application.

关 键 词：匀相法 N-乙酰-Β-D-氨基葡萄糖苷酶 6-甲基-2-硫代吡啶-N-乙酰-β-D-氨基葡萄糖苷 

分 类 号：Q555.4[生物学—生物化学]