CAJ | 학술논문

人们研究并逐步认识了纳米金与DNA等相互作用的特性和条件,并对以纳米金为基础发展起来的生物条码技术进行了更加深入的研究与应用.基于纳米金的生物条码及其相关技术研究与应用日益广泛,并不断有创新,本研究应用纳米技术并结合分子探针技术对低浓度的抗原水平进行检测.该生物条码技术常被用于检测某些蛋白、核酸、细胞或细胞因子的水平,并由此形成了不同于PCR、ELISA但仍具有高灵敏度特性的检测方法.通过磁性微球标记单抗以捕获、富集待测抗原,修饰多抗与亲和素的纳米金并结合分子信标探针起到信号扩增的作用.通过二硫键(-S-S-)分子信标探针茎部3'端的碱基T连接一生物素分子.待测抗原可将磁球与纳米金系统进行连接,然后应用二硫苏糖醇(DTT)将连接于纳米金上的MB切割下来,并与其环部完全互补配对的靶序列杂交后测定荧光强度.荧光强度与待测抗原水平(含量)相关.
인문연구병축보인식료납미금여DNA등상호작용적특성화조건,병대이납미금위기출발전기래적생물조마기술진행료경가심입적연구여응용.기우납미금적생물조마급기상관기술연구여응용일익엄범,병불단유창신,본연구응용납미기술병결합분자탐침기술대저농도적항원수평진행검측.해생물조마기술상피용우검측모사단백、핵산、세포혹세포인자적수평,병유차형성료불동우PCR、ELISA단잉구유고령민도특성적검측방법.통과자성미구표기단항이포획、부집대측항원,수식다항여친화소적납미금병결합분자신표탐침기도신호확증적작용.통과이류건(-S-S-)분자신표탐침경부3'단적감기T련접일생물소분자.대측항원가장자구여납미금계통진행련접,연후응용이류소당순(DTT)장련접우납미금상적MB절할하래,병여기배부완전호보배대적파서렬잡교후측정형광강도.형광강도여대측항원수평(함량)상관.
With better understanding of the characteristics and conditions of the interaction between the gold nanoparticles and DNA,people are developing the bio-bar-code growth technology based on the gold nanoparticles.The method is widely applied to detect the level or microconcentration of some proteins,nucleic acid,cells and cytokines.New methods are coming into being with higher sensitivity as distinct from the PCR and ELISA.The biobar-code and the related technology based upon the gold nanoparticles are being continuously innovated.In this paper,the microconcentration of the antigen IgG is detected by applying nanotechnology and molecular probes technology in the experiment.The magnetic microbeads modified with the monocolon antibody are used to capture the target antigen IgG,to modify the gold nanoparticles with the polycolonal antibody and the avidin combined with the molecular beacons,thus to modify the biotin through the disulfide bonds at the T base position of the 3' terminal of the stem,and to amplify the signal in cascades.The magnetic microbeads can be combined with the gold nanoparticles through the detected antigen IgG.Then the DTT could cut the MB off the gold nanoparticles through the reduction effect and the free MB can be hybridized with its target sequence.Then the fluorescence density is measured with Spectrofluorometer.The results show that there are some relationships between the fluorescence density and the concentration of the antigen lgG.The detection sensitivity of the research is very high,which reaches 1pg/ml,about the 6.67 fM.