血清精氨酸代琥珀酸裂解酶速率测定法的建立及初步临床应用

Establishment of enzymatic kinetic assay of serum argininosuccinate lyase activity and its preliminary clinical application

目的建立适用于自动生化分析仪的血清精氨酸代琥珀酸裂解酶（ASL）活性速率测定法，并进行方法学评价和初步临床研究。方法根据ASL催化的化学反应，以及自动生化分析仪工作特点，建立特异性的偶联酶促反应体系，并对建立的方法进行方法学评价。共测定309例肝病患者、269例非肝病患者和40名健康人血清ASL和传统的肝病酶学指标ALT、AST活性。结果建立了一种新的可在全自动生化分析仪上测定血清ASL活性的动力学方法。方法学评价研究表明，本法的批内变异系数均值为4．0％，天问变异系数均值为5．9％，平均回收率是100．5％，在0—167．7U／L间有良好的线性范围，最低检测限为0U／L。干扰试验提示：胆红素〈342μmol／L、常用抗凝剂在抗凝浓度内不会产生干扰，Hb〉0．06g／L时产生明显干扰。初步临床分析显示非肝病患者血清ASL水平与对照组问差异无统计学意义（q=0．027，P=0．979），而与ATL和AST间差异均有统计学意义（ALT：q=6．461，P=0．000；AST：q=6．481，P=0．000）。结论成功建立了适用于自动生化分析仪完成的ASL活性速率测定法，其有可能是一个较好的肝病实验诊断指标。

Objective To establish a continuous monitoring assay of serum argininosuccinate lyase （ASL） activity with automatic biochemistry analyzer, and perform methodology validation and preliminary clinical application. Methods According to the chemical reaction catalyzed by ASL and the working characteristics of automatic biochemistry analyzer, an enzyme coupled reaction system with high specificity was set up, and the methodology validation was performed. Three hundred and nine patients with various liver diseases, 269 non-liver disease patients and 40 healthy controls were enrolled in this study. Serum ASL, ALT, and AST level were determined in all subjects. Results A new kinetics assay of ASL activity was set up with automatic biochemistry analyzer. The methodological validation demonstrated that inter-assay and intra-assay coefficient of variation were 4. 0% and 5. 9% respectively and the mean recovery was 100. 5%. The linear range was 0-167.7 U/L. The lowest detection limit was approximately 0 U/L. The interference test showed that there is no significant interferences while the concentration of bilirubin is less than 342 μmol/L or commonly used anticoagulants is employed at their routine concentrations. However, interference was significant when Hb level is more than 0. 06 g/L. Preliminary study of clinical application showed that there was no significant difference of serum ASL level between non-liver disease group and healthy group （q = 0. 027, P = 0. 979 ）, but there was significant differences for both serum ALT and AST levels （ALT：q=6.461,P=0.000;AST：q=6.481,P=0.000 ）. Conclusions A continuous monitoring assay for the determination of serum ASL activity is successfully established. Serum ASL may be a good biomarker for liver injury.