利用生物条形码技术对相思子毒素进行微量检测  

Bio-bar Codes Assay(BCA) in Ultrasensitive Detection of Abrin

作　　者：张立营[1] 冯玉奎[1] 孙英姿[1] 孙宇[2] 高波[1] 

机构地区：[1]济南军区青岛第二疗养院检验科,山东青岛266071 [2]北京305医院检验科,北京100050

出　　处：《中国误诊学杂志》2010年第16期3784-3786,共3页Chinese Journal of Misdiagnostics

基　　金：青岛市公共领域科技支撑计划项目(编号:09-1-1-28-nsh)

摘　　要：目的建立相思子毒素的高灵敏检测方法。方法利用相思子毒素的多抗及特异DNA链标记的金纳米颗粒NP探针和相思子毒素单抗标记的磁性微球MMP探针,形成MMP-相思子毒素-NP三明治复合物后再利用去杂交将NP探针上标记的DNA链释放出来,通过PCR方法或芯片检测方法鉴定这些释放的DNA链就可确定相思子毒素的存在。结果建立了相思子毒素的生物条形码检测体系,检测灵敏度可达1 fg/ml。和临床上常规的检测方法相比,其检测灵敏度可达常规ELISA的106倍。结论本资料为发展高灵敏度的相思子毒素的生物条形码检测试剂盒鉴定了基础。Objective To establish a highly sensitive method in detecting abrin.Method NPs（nanoparticle probes） that were encoded with DNA was unique to Abrin and polyclonal antibodies agaist abrin and MMPs（magnetic microparticle probes） with monoclonal antibodies that specifically bind Abrin were prepared.After forming MMPAbrin-NP sandwich complex,dehybridization of the oligonucleotides on the nanoparticle probe surface allowed the determination of the presence of Abrin by identifying the oligonucleotide sequence released from the nanoparticle probe through PCR or chip-based detection.Result The Bio-bar Codes Assay system for detecting Abrin was established,and the sensitivity limit was about 1fg/mL,Comparable clinically accepted conventional ELISA assays for detecting the same targe,six orders of magnitude less sensitive than what was observed with this method.Conclusion This study lays a foundation for establishing highly sensitive Abrin Bio-bar Codes Assay kit.

关 键 词：鸡骨草/化学 毒素类 生物学/分析 自动数据处理 生物学鉴定法 

分 类 号：R284.1[医药卫生—中药学]