摘要
目的:建立微孔板化学发光法检测5'-核苷酸酶(5'-NT)新技术。方法:5'-核苷酸酶经嘌呤核苷磷酸化酶-黄嘌呤氧化酶-尿酸酶-过氧化物酶(PNP-XTO-UAO-POD)偶联途径反应产生过氧化氢H2O2,H2O2在过氧化物酶(POD)的作用下与鲁米诺体系反应发光,与标准品比较后,计算出5'-NT的活性。结果:该法线性范围为0.02 U/L~120.8 U/L,平均回收率为98.8%,批内相对标准差(RSD)和批间相对标准差分别为2.2%~3.9%和3.5%~4.9%,与参考方法的线性回归方程和相关系数分别为Y=0.978X+0.084,r=0.9962。结论:用PNP-XTO-UAO-POD偶联化学发光分析法测定血清血清5'-NT,方法简便、快速,灵敏可靠,适合临床推广应用。
Objective:To establish a microperforated panel chemiluminescence method for 5' - NT analysis in serum. Methods: 5' - nucleotidase (5' - NT) hydrolyzed inosine 5' - monophosphate ( 5' - IMP) to generate inosine, then the inosine was converted to H2 02 by purine nueleoside phosphorylase - xanthine oxidase - uricase - peroxidase ( PNP - XTO - UAO - POD) - coupled reaction pathway. The H2 02 reacted with luminol system and produced chemical 1 , then the 5' - NT activ- ity can be ca/culated by eomparation with the standard 5' - NT activity. Results: The method was linear in the range of 0.02 U/L - 120.8 U/L, the average rate of recovery was 98.8% , the within - run and between - run RSDs were 2.2% ~ 3.9% and 3.5% -4.9% respectively. Comparing this method with the reference method, the regression equation was Y =0. 978X +0.084, r = 0. 9962. Conclusion: This method is simple, sensitive and suitable for clinical application.
出处
《中国卫生检验杂志》
北大核心
2012年第1期28-30,共3页
Chinese Journal of Health Laboratory Technology
基金
浙江省科技厅分析测试科技计划项目(2008F70051)