| | | | | 蛋白质芯片定量检测乙肝表面抗原系统的建立 | | | 吴昊;罗进;郭晏海;王小明;闫小君;李丁 | | | 目的:建立以蛋白质芯片检测乙肝表面抗原的方法,以期对乙肝患者血清中HBsAg进行定量检测,帮助动态分析病情和评估疗效。方法:用点样仪将一种抗-HBsAg单抗点到硝酸纤维素膜(NC膜)上制成芯片,HRP标记另一种抗-HBsAg单抗,以双抗夹心法捕获血清中HBsAg,建立HBsAg浓度与检测点灰度间关系的标准曲线并通过芯片阅读仪进行定量分析。结果:建立的蛋白质芯片检测系统能快速准确地对血清中HBsAg进行定性和定量检测,系统的定量检测范围是1~10000 ng/ml。结论:蛋白质芯片检测系统具有微型化和自动化的优点,与PCR HBV-DNA相比其操作更简便,所需时间更短,可以代替传统的ELISA方法,是更为科学的疗效评估方法。
【作者单位】：第四军医大学药学系药物基因组学教研室全军基因诊断技术研究所国家肿瘤生物重点实验室 西安710032(吴昊;罗进;郭晏海;闫小君;李丁);陕西省生物诊断与治疗工程技术研究中心 西安710032(王小明)
【关键词】：蛋白质芯片;乙型肝炎表面抗原(HBsAg);定量检测;硝酸纤维素膜
【分类号】：R446.6
【DOI】：CNKI:SUN:ZWJZ.0.2008-05-040
【正文快照】：
蛋白质芯片技术是指将蛋白质或多肽固化于固相支持物表面,通过蛋白质-蛋白质相互作用,对样品中靶蛋白分子进行高通量检测的技术[1]。目前已经广泛的被应用于临床和科研。乙型肝炎表面抗原(HBsAg)是乙型肝炎病毒(HBV)外壳的一种结构成分,为HBV感染的标志物之一,存在于患者血清小 | | |
| | |  推荐 CAJ下载  PDF下载 | | | CAJViewer7.0阅读器支持所有CNKI文件格式，AdobeReader仅支持PDF格式 | | | | Establishment of protein microarrays quantitative detect system for hepatitis B virus surface antigen | | | Wu Hao1;Luo Jin1;Guo Yan-Hai1;Wang Xiao-Ming2;Yan Xiao-Jun1;Li Ding1(1.PLA Institute of Genetic Diagnosis;State Key Laboratory of Cancer Biology;Department of Pharmacogenomics;School of Pharmacy;Fourth Military Medical University;Xi′an 710032;China;2.Shaanxi Province Biologic Diagnosis and Therapy Engineering Research Center;Xi′an 710032;China) | | | Objective:To establish a novel method of quantitatively detecting HBsAg in patients serum by protein microarrays to analyze patients′ state and evaluate curative effect.Methods:Using microarrayer to print a kind of anti-HBsAg antibody on nitrocellulose membrane(NC membrane) to make the Microarrays.Another anti-HBsAg antibody was marked by HRP.The HBsAg in serum was detected by microarray with double antibody sandwich method.To establish the HBsAg concentration standard curve measured by chroma.And measuring HBsAg concentration in serum.Results:The protein microarrays detection system can quickly and accurately detect HBsAg qualitatively and quantitatively in serum.The range of the assay was 1~10000 ng/ml HBsAg.Conclusion:Our protein microarrays detect system has advantage of the miniaturization and automation.The system operates more simply and quickly than the method of PCR HBV-DNA and ELISA.The system can replace the traditional method of ELISA to detect HBsAg.It is more scientific method of evaluating curative effect.
【Keyword】：Protein microarrays;Hepatitis B virus surface antigen;Quantitative detection;Nitrocellulose membrane |
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