生物素-亲和素-DNA纳米颗粒信号放大检测载体的构建  

Construction of Biotin-Streptavidin-DNA Nanoparticle Signal Amplification Carrier for Nucleic Acid Assay

作　　者：王海河[1] 赵春燕[1] 李凡[1] 

机构地区：[1]吉林大学白求恩医学院病原生物学教研室,长春130021

出　　处：《中国生物制品学杂志》2009年第1期27-30,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金资助课题(30672007)

摘　　要：目的构建用于检测细菌、病毒等病原体DNA的生物素-亲和素纳米颗粒信号放大载体。方法设计并合成两端和一端标记生物素的寡核苷酸探针(2B/1B-DNA),与抗生蛋白链菌素葡聚糖(Poly-STV)通过生物素与亲和素作用,偶联形成大分子纳米网状颗粒,并筛选最佳探针类型及其长度、2B-DNA和Poly-STV的浓度及比例和反应条件。结果2B-DNA和Poly-STV形成纳米颗粒信号放大载体的最佳浓度分别为1μmol/L和5μmol/L,最佳浓度比为1∶5,2B-DNA长度为60bp,缓冲液为Buffer B,反应条件为55℃,800r/min,20min。结论已成功构建了生物素-亲和素大分子纳米颗粒,可作为DNA检测信号放大技术的备选载体。Objective To construct a biotin-streptavidin-DNA nanoparticle signal amplification carrier for assay of DNA in pathogens such as virus and bacterium. Methods Mono- and bis-biotinylated oligonucleotide probes （1B/2B-DNA） were designed and synthesized, then coupled to streptavidin-labeled dextran （Poly-STV） through the interaction of biotin and streptavidin to construct a macromolecular nanoparticle signal amplification carrier. The kind and length of probe, the concentrations of 2B-DNA and Poly- STV, the ratio of 2B-DNA to Poly-STV as well as reaction condition were optimized. Results The optimal concentrations of 2B-DNA and Poly-STV for construction of the carrier were 1 and 5 μmol / L respectively, the optimal ratio of 2B-DNA to Poly-STV was 1 ： 5, the optimal length of 2B-DNA was 60 bp, the optimal reaction condition was 55℃, 800 r/min for 20 min. Conclusion The biotin- streptavidin-macromolecular nanoparticles were successfully constructed, which might be used as a candidate cartier for DNA signal amplification detection technology.

关 键 词：生物素 亲和素 纳米颗粒 信号放大 载体 

分 类 号：Q789[生物学—分子生物学]