景晟（上海复星长征医学科学有限公司 上海 200444）

自制5’-核苷酸酶试剂在生化检验中的应用

景晟*
（上海复星长征医学科学有限公司 上海 200444）

摘 要目的：评价自主研发的5’-核苷酸酶试剂（速率法）在自动化检测血清或血浆中5’-核苷酸酶活性的应用。方法：采用奥林巴斯AU680全自动生化分析仪，通过重复性、线性试验、测定血清或血浆中5’-核苷酸酶活性，进行方法学评价并与美国Diazyme 5’-核苷酸酶试剂（速率法）比较。结果：两个水平质控中酶活性的变异系数（CV）分别为2.83% 和1.96%，在 5～200 U/L酶活范围内，线性良好。两种试剂测定血清5’-核苷酸酶活性的相关系数较好，相关系数r 为0.995。结论：自主研发的5’- 核苷酸酶试剂盒操作简便、快速、准确、稳定，结果准确，可满足大批量临床样本的常规和自动化分析，是一种值得推广的方法。

关键词5’-核苷酸酶 诊断试剂 临床研究

The application of the homemade 5’-nucleotidase reagent in biochemical analysis

JING Sheng*
(Shanghai Fosun Long March Medical Science Co. Ltd., Shanghai 200444,China)

ABSTRACTObjective: To evaluate the application of the independent innovation reagent in the automatic determination of 5’-nucleotidase (5’-NT) activity in the clinical serum or plasma samples. Methods: 5’-NT activities were determined with Olympus AU680 automatic biochemical analyzer by repeatability and linearity tests, the evaluation of methodology was carried out and the results were compared with those determined by the reagent of Diazyme Laboratories. Results: The coefficients of variation (CV) of the enzyme activity at two levels of quality control were 2.83% and 1.96%. The determination of enzyme activity showed a good linearity over the range of 5~200 U/L. There was a good correlation between two reagents (r=0.995). Conclusion: The 5’-nucleotidase reagent made by our company can provide a simple, quick, accurate and stable method for the determination of 5′-NT activity in the clinical serum or plasma samples and meet the requirement of the automated analysis of mass clinical samples. This method may be worthy of promotion.

KEY WORDS5’-nucleotidase; clinical research; diagnostic reagent

1934年Reis第一次描述了心脏和骨骼肌中5′-核苷酸酶（EC3.1.3.5）的活性[1]。它的主要功能是将核苷酸5′-C上连接的磷酸催化水解生成核苷和正磷酸，其在细胞生长发育、运动、纤维蛋白合成、神经传递、淋巴细胞的黏附、再循环、免疫应答等方面发挥重要作用及在缺血性心脏病、缺血性肾脏病、白血病、肿瘤等疾病的发生和发展过程中的作用越来越受到人们的重视[2-4]。目前测定5′-核苷酸酶活性的方法有很多，包括钼蓝显色法[5]、Kalckar 氏法[6]、改进的Kalckar 氏法[7]、酶法[8]和速率法[5]。其中钼蓝显色法和速率法是最常用的方法，但是钼蓝显色法所需试剂种类多，操作繁琐、费时，不适宜自动化分析；……