新型肝损伤血清标志物Pep5定量检测方法的建立及临床应用  

Development and application of quantitate assay for serum biomarker polypeptide 5 of liver injury

作　　者：傅海媛[1,2,3] 侯利平[2] 周晓明[2] 黄亚娟[2] 宋纯艳[2] 孙云波[2] 汪洋[2] 杨保安 肖汉族 甄蓓[2] 魏开华[2,3] 王海滨[6] 

机构地区：[1]解放军总医院,北京100853 [2]军事医学科学院放射与辐射医学研究所,北京蛋白质组研究中心,蛋白质组学国家重点实验室,国家蛋白质科学中心(北京) [3]安徽省芜湖市第二人民医院,北京102206 [4]北京正旦国际科技有限责任公司,241000 [5]湖南金健药业有限责任公司,北京102206 [6]解放军三○二医院,常德415001

出　　处：《检验医学与临床》2013年第20期2641-2643,共3页Laboratory Medicine and Clinic

基　　金：国家高技术研究发展计划863项目(2012AA020205);卫生部传染病重大专项(2008ZX10002-016)

摘　　要：目的建立定量检测血清多肽Pep5的酶联免疫吸附方法(ELISA),用于判定肝损伤进程的研究。方法以待测血清样本稀释液包被酶联板,5%脱脂奶粉封闭,辣根过氧化物酶标记的抗Pep5单克隆抗体作为标记抗体,建立定量检测人血清多肽Pep5的ELISA,并用于临床检测690例肝损伤及健康血清样本。结果灵敏度为1.25ng/mL;线性范围(0～24ng/mL)内相关系数为0.998 7;与乙酰胆碱酯酶、层黏连蛋白、透明质酸、三型前胶原交叉反应率小于1.00%;添加回收率在99.09%～107.08%;批内、批间变异系数(CV)分别为5.62%～7.32%、6.21%～6.45%。与其他肝损伤指标乙酰胆碱酯酶试剂盒进行对比试验,表明两种检测方法有一定的相关性(r=0.68)。临床检测结果表明血清多肽Pep5水平随着肝损伤程度加重而升高,与正常血清差异有统计学意义(P<0.01)。结论多肽Pep5可用于肝损伤进程定量血清学检测,检测方法简便、可靠,为后续深入研究多肽Pep5的功能及相关诊断试剂盒的研制和开发提供了科学依据。Objective To establish an enzyme-linked immunosorbent assay（ELISA） for polypeptide5（Pep5） in human serum and apply it to the serological diagnosis of the progression of liver injury .Methods Microplate was precoated with sample diluents ,blocked with 5% milk ,and samples were detected with anti Pep5 McAb labled with HRP .And 690 clinical samples were detected by method metioned above .Results Sensitivity was 1 .25 ng/mL .Line-ar measurement range was 0-24 ng/mL .Recovery rate was 99 .09% -107 .08% .Intra-and inter-assay coefficinet of variation were 7 .32% and 6 .45% respectively ,or less .The cross-reaction rate with acetylcholinesterase ,laminin ,hy-aluronic and procollagen were all less than 1 .00% .Clinical detecting showed that levels of Pep5 increased with the exacerbation of liver injury （P〈0 .01） .Compared with acetylcholinesterase kit ,the relative coefficient （r） was 0 .68 . Conclusion Pep5 could be used for the quantitative serological tests for the progression of liver injury ,which might be reliable ,simple and convenient ,and could provide scientific evidences for the further study of Pep5 function and de-velopment of related diagnostic kits .

关 键 词：多肽Pep5 酶联免疫吸附法 肝损伤 血清学诊断 

分 类 号：R657.3[医药卫生—外科学]