摘要
目的 建立对杨磷酶（PON-1）自动化分析方法.方法 以对硝基苯磷酸二乙酯（对氧磷,paraoxon）为底物,在甘氨酸-氢氧化钠（Glycine-NaoH）缓冲液系统下生成色源对硝基酚.通过测定对硝基酚在405 nm处的吸光度变化率,可计算出PON-1的活性.结果 最适pH值为10.2,最适底物浓度为4.2 mmol/L,Ca2＋浓度为2.5 mmol/L,米氏常数（Km）为0.4 mmol/L.批内、批间变异（CV%）分别为1.07%和2.20%.健康对照组血清PON-1活性为（422.1±78.2）U/L;非冠心病组和冠心病组PON-1活性分别为（411.9±66.9）U/L和（284.1±48.3）U/L,冠心病组PON-1活性显著低于非冠心病组,差异具有统计学意义（P〈0.01）,非冠心病组和健康对照组检测结果无显著差异.结论 该法简便、快捷、精确,可用于自动化分析,可用于心血管疾病诊断的诊断及疗效观察.
Objective To establish a kinetic monitoring assay for detecting PON.Methods Nitrobenzene diethyl phosphate was used as substrate.PON-1 could catalyze the substrate to produced chromogenic p-nitropheny1 in Glycine-NaoH buffer.The activity of PON-1 could be obtained through determining the product at 405nm.Results The optimum pH was 10.2 and substrate concentration was 4.2 mmol/L.Ca2＋ concentration was 0.25 mmol/L.Km was 0.4mmol/L.Intra-assay CV value was 1.07% and inter-assay CV value was 2.20%.The activity of PON-1 in normal control group was（422.1±78.2）U/L.The activity of PON-1 in Non coronary heart and coronary heart group were（411.9±103.9）U/L and（284.1±78.3）U/L.The activity in coronary heart groupIn was lower than the non coronary heart group,there was a significant difference（P〈0.01） Between two groups.There was no significant difference bewteen non coronary heart group and normal control group.Conclusion The method was simple,rapid,accurate,and suitable for automatic analysis of PON-1.It can be used in the diagnosis and treatment of cardiovascular diseases.
作者
王蓉
沈安达
张建华
WANG Ronga SHEN An-da ZHANG J ian-hua(Department of Clinical Laboratory, the Second People＇s Hospital of Shanghai , Shanghai 200011, China Department of Chemistry, Shanghai J iaotong University School of Medicine, Shanghai 200025, China)
出处
《检验医学与临床》
CAS
2017年第A01期14-17,共4页
Laboratory Medicine and Clinic
基金
上海市黄浦区优秀学科带头人资助项目.
关键词
对杨磷酶
动力学法
冠心病
冠脉造影
paraoxonase
kinetic method
coronary heart disease
coronary angiography
