5-羟基吲哚乙酸均相酶免疫法的建立  

The Establishment of Homogeneous Enzyme Immunoassay for 5-Hydroxyindoleacetic Acid

作　　者：魏雪梅 冯杰[1] 梁辰[1] 邓子辉[1] 薛辉[1] 陆丽华 何秀丽 虞留明 高艳红[1] 颜光涛[1] WEI Xuemei;FENG Jie;LIANG Chen;DENG Zihui;XUE Hui;LU Lihua;HE Xiuli;YU Liuming;GAO Yanhong;YAN Guangtao(Department of Clinical Laboratory Center,the First Medical Center of the PLA General Hospital,Beijing 100853,China;Suzhou Evermed Biomedical Co.,Ltd,Suzhou 215163,China)

机构地区：[1]解放军总医院第一医学中心医学检验中心,北京100853 [2]苏州博源医疗科技有限公司,江苏苏州215163

出　　处：《标记免疫分析与临床》2020年第10期1741-1744,1749,共5页Labeled Immunoassays and Clinical Medicine

基　　金：国家重点研发计划课题子课题(编号:2017YFF0205401)。

摘　　要：目的采用均相酶免疫法检测5-羟基吲哚乙酸(5-HIAA)的浓度。方法样本中游离的5-HIAA与葡萄糖六磷酸脱氢酶-5-羟基吲哚乙酸偶联物竞争性结合抗体位点,游离出来的5-羟基吲哚乙酸酶标偶联物催化NAD+转化为NADH,样本中的5-HIAA浓度与NADH的生成量成正比,通过340 nm吸光值的变化即可计算出5-HIAA的含量。结果据本研究建立的方法,健康人5-HIAA参考范围为0~8.0 mg/L,线性范围为2.0~98.0 mg/L,准确度相对偏倚<5%,批内变异<5%,批间变异<5%,携带污染率和试剂上机21d稳定性等符合判断标准。结论该5-HIAA均相酶免疫法检测的试剂盒性能满足临床检测要求,可以推广使用。Objective To detect the concentration of 5-hydroxyindoleacetic acid(5-HIAA)by homogeneous enzyme immunoassay.Methods Free 5-HIAA and glucose hexaphosphate dehydrogenase-5-hydroxyindoleacetic acid conjugate could compete for binding to antibody site,then free 5-hydroxyindoleacetic acid enzyme conjugate catalyzed NAD+conversion to NADH.The concentration of 5-HIAA in the sample was directly proportional to the production of NADH.The content of 5-HIAA could then be calculated by changing the absorbance value at 340 nm.Results According to the method established in this study,the reference range of healthy people was 0-8.0 mg/L,the linear range was 2.0-98.0 mg/L,the relative bias of accuracy was less than 5%,intra assay variation was less than 5%,and inter assay variation was less than 5%.The carrying contamination rate and the 21-day stability of reagents met the judgment criteria.Conclusion The performance of the 5-HIAA homogeneous enzyme immunoassay kit meets the requirements of clinical detection and can be widely used clinically.

关 键 词：5-HIAA 均相酶免疫法 类癌综合征 

分 类 号：R392-33[医药卫生—免疫学]