纳米磁性颗粒标记的免疫层析法定量检测人乙型肝炎表面抗原  

Development of a Rapid Quantitative Magnetic Nanoparticle-Based Immunochromatographic Test for Human Hepatitis B Surface Antigen

作　　者：孙建斌[1] 王伟华[1] 雷小英[1] 郭晏海[1] 颜真[1] 杨静华 

机构地区：[1]第四军医大学药学院药物基因组学教研室 [2]Department of Surgery,Boston Veterans Af-fairs Healthcare System,Boston University School of Medicine,Boston 02215,Massachusetts,USA

出　　处：《生物技术通讯》2012年第1期94-97,共4页Letters in Biotechnology

基　　金：国家高技术研究发展计划(2007AA022004);"十一五"国家"艾滋病和病毒性肝炎等重大传染病防治"科技重大专项(2009ZX10004-711)

摘　　要：目的:应用纳米磁性颗粒标记的免疫层析法,研制可应用于乙肝表面抗原(HBsAg)快速定量检测的层析试纸条。方法:用1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺(EDC)/N-羟基琥珀酰亚胺(NHS)交联的方法标记纳米磁珠,喷膜仪喷点硝酸纤维膜;根据双抗体夹心法原理建立免疫层析试纸条,对HBsAg特异性抗体捕获的磁信号进行检测,并对磁信号检测结果进行统计学分析和评价。结果:建立了HBsAg纳米磁性免疫层析试纸条,最低限度检测为0.1 ng/mL的HBsAg抗原,检测灵敏度达到了同类产品ELISA分析法的标准,且检测时间控制在5 min内;经检测临床血清标本证实,该方法可根据磁信号定量检测乙肝患者血清中HBsAg的浓度。结论:HBsAg纳米磁性免疫层析方法具有简单快速、灵敏度高的特点,可应用于临床血清样本中HBsAg的检测;该方法为体内极微量抗原抗体的快速检测建立了新模式。Objective： Magnetic nanoparticle was labeled to establish an immunochromatography strip for the quantitative detection of HBsAg in patients serum rapidly.Methods： Magnetic nanoparticles were catalyzed by ethyl（dimethylaminopropyl） carbodiimide（EDC）/N-Hydroxysuccinimide（NHS）.Antibodies were sprayed onto the NC membrane by Biodot biojet to establish an immunoassay test strip in sandwish assay,which could arrest the magnetic signals.Then the results of magnetic signals were analyzed and evaluated.Results： A magnetic immunoassay strip for the detection of HBsAg in patients serum was established.The sensitivity of this method could reach 1 ng/mL,equivalent to the ELISA method.And one single test could be completed in 5 min.Conclusion： It is a simple,rapid,high sensitivity and quantitative method for detecting not only HBsAg,but a trace amount of antigen/antibody in patients samples during clinical application.

关 键 词：磁性纳米颗粒 免疫层析 定量 乙肝表面抗原 快速检测 

分 类 号：R392.1[医药卫生—免疫学] Q502[医药卫生—基础医学]