丙型肝炎病毒分片段抗体检测蛋白质芯片质控参比品的建立及质量验证  

Establish and validation of reference for quality control of protein-chip for different HCV antibody detection

作　　者：张文[1,2] 周枚芬 陈立炎 丁亚平 黄坚 耿永尧 王升启[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]深圳益生堂生物企业有限公司,深圳518033

出　　处：《生物技术通讯》2004年第3期248-250,共3页Letters in Biotechnology

基　　金：国家自然科学基金重点项目(39889001);军队杰出人才基金项目

摘　　要：建立能同时检测丙型肝炎病毒(HCV)嵌合抗体、核心抗体和NS3、NS4、NS5抗体的蛋白质芯片质控参比品,对质控合格的芯片进行质量验证。用3种HCVEIA试剂分别检测从3家医院收集的丙型肝炎病毒感染患者血清及其他非HCV感染患者血清,从3种EIA试剂同时阴性或阳性的血样中挑取阳性和阴性血清,然后用RNAhybPCR试剂进行检测,从中再选取部分样本用RIBA3.0进行检测,确定HCV分片段抗体检测蛋白质芯片质控参比品。经质检合格的芯片用中国药品生物制品检定所的HCV参比品进行检定。通过490例临床标本的检测对芯片的质量进行进一步的验证。从收集的240份丙型肝炎病毒感染患者血清及其他非HCV感染患者血清筛选出30份血样(15份阳性,15份阴性)作为HCV分片段抗体检测蛋白芯片质控参比品。中国药品生物制品检定所的80份HCV参比品检定结果表明,混合抗体阳性检出率为39/40,阴性符合率为40/40,总符合率为98.7%;核心抗体阳性检出率为27/40,阴性符合率为40/40;NS3抗体阳性检出率为26/40,阴性符合率为39/40;NS4抗体阳性检出率为19/40,阴性符合率为40/40;NS5抗体阳性检出率2/40,阴性符合率为40/40。490例临床标本的检测结果表明,对于194例HCV阳性标本,蛋白质芯片混合抗体与ELISA的符合率达99.5%,分片段抗体符合率达97.4%,两种方法检测?Serum of patients infected by HCV and without infected by HCV collected from three hospital were detected by three HCV EIA reagents.Positive and negative serum validated by all the three reagents were chose respectively and then detected by RNAhyb PCR reagents.Furthermore,after detected by RIBA reagents,parts of them was achieved as reference for the quality control of the proteinchip.The qualified proteinchip was identified by HCV standard of National Institute for the Pharmaceutical and Biology Products(NICPBP).The quality of the proteinchip was further evaluated through detec-tion of490clinical samples.30serum with15positive and15negative were achieved from240collected serums of pa-tients infected by HCV and considered as reference of the quality control of the proteinchip for the detection of different HCV antibody.Through detected with80standard of NICPBP,The results shows the positive and negative conformity ra-tios were39/40and40/40,27/40and40/40,26/40and39/40,19/40and40/40,2/40and40/40for combined anti-body,core antibody,antibody to NS3,antibody to NS4,antibody to NS5respectively compared with ELISA.The results of clinical sample show that the conformity ratio of combined antibody and different antibody of the proteinchip were99.5%and97.1%respectively compared with ELISA.The unconformity samples were validated by RIBA reagents and the result shows the conformity of proteinchip with RIBA reagents was high consist.The results of296clinical samples without HCV infected show negative all.The prepared reference for quality control of protein-chip for the detection of different HCV antibody can be used as quality control for the production of the proteinchip.The produced proteinchip can pass the demand of standard of NICPBP and can be used in clinical detection.

关 键 词：丙型肝炎病毒 蛋白质芯片 质量验证 生物制品 抗体 检测 

分 类 号：R512.63[医药卫生—内科学]