本文主要研究内容
作者李政(2019)在《基于量子点—适配体的荧光传感器用于检测癌症标志物MUC1》一文中研究指出:现阶段,癌症是人类健康的一大威胁,癌症的治愈也是科研工作者需要攻克的难关,癌症病发的早期阶段是进行介入治疗的最佳时期。癌症相关蛋白是一类可以衡量疾病当前和未来状态的指标分子,精准检测出其在患者体内的含量则有助于医生对患者进行最有效的临床诊断和介入治疗,因此,对癌症相关蛋白的灵敏检测显得尤为重要。MUC1是一种位于上皮细胞顶端膜部位的糖蛋白,会在已丧失极性的癌细胞表面过量表达,因此,其在人体内的含量可以作为癌症早期诊断的指标。然而在细胞癌变的早期阶段,MUC1的含量较低,不易检测,有待探索简便、高效、灵敏的检测方法。量子点是最常用的荧光探针之一,宽激发、窄发射、尺寸可调以及高量子产率的特点使得量子点在不同研究领域得到了广泛应用,例如:生物成像、光学编码、生物传感等。适配体是指对目标分子有特异性亲和力的寡核苷酸或者肽段,常被设计为检测探针用于目标物的特异性灵敏检测。基于适配体与MUC1的特异性结合作用,可将适配体设计为检测探针用于MUC1的灵敏检测。磁性微球也可简称为“磁珠”,具有高比表面积、易修饰、易分离的特点,广泛的应用于核酸、蛋白质、小分子物质以及癌细胞等方面的检测。杂交链式反应(HCR)是一种常用的信号放大技术,在引发链的作用下,发卡结构的DNA序列H1和H2可自组装杂交成一条长的DNA双链,在无任何核酸酶的参与下,仅通过DNA级联杂交反应即可实现检测信号扩增。本论文基于适配体对蛋白特异性识别作用并结合杂交链式反应(HCR)信号放大技术,且利用磁珠易分离的特性显著降低背景干扰,设计了两种量子点荧光探针,实现了对MUC1的简便、灵敏及特异性的检测,主要研究内容如下:(1)参照本课题组已有的方法,制备了CdZnTeS量子点;同时在检测体系中引入核酸“分子光开关”钌(Ⅱ)配合物—其在水溶液中基本无荧光信号,当嵌入DNA双链中时,可以测得明显的荧光信号;利用钌(Ⅱ)配合物对量子点的荧光猝灭作用及钌(Ⅱ)配合物嵌入DNA双链时,对量子点无猝灭作用的特性,将可特异性识别MUC1的适配体设计为发卡结构靶向MUC1,启动杂交链式反应(HCR)将信号放大,经过磁分离后测定上清液的荧光强度。据此,可以根据钌(Ⅱ)配合物对CdZnTeS量子点的猝灭程度来定量检测MUC1,线性范围0.2-100 ng/mL(LOD=0.13 ng/mL,S/N=3),此方法也可对血清中MUC1含量进行灵敏测定。(2)将前一章检测MUC1的方法进一步改进,无需引入钌(Ⅱ)配合物。采用一锅法直接合成DNA功能化量子点,DNA功能化量子点通过与互补单链杂交引发杂交链式反应形成长的双链。当体系中存在MUC1时,此前保持稳定发卡结构的H0由于与MUC1的折叠缠绕,结构发生改变,暴露出单链末端,并可以与发卡结构的Fuel发生互补杂交,进而使Fuel尾端成为直链结构,从而可以与大量的S1和S2(DNA-CdTe:Zn2+QDs)发生周期的互补杂交,生成杂交链式反应产物(双链中含有大量的量子点)。经过磁分离后,直接检测上清液的荧光强度,即可对MUC1进行定量检测,线性范围0.15-100 ng/mL(LOD=0.11ng/mL,S/N=3),并可用于血清中MUC1的灵敏检测。
Abstract
xian jie duan ,ai zheng shi ren lei jian kang de yi da wei xie ,ai zheng de zhi yu ye shi ke yan gong zuo zhe xu yao gong ke de nan guan ,ai zheng bing fa de zao ji jie duan shi jin hang jie ru zhi liao de zui jia shi ji 。ai zheng xiang guan dan bai shi yi lei ke yi heng liang ji bing dang qian he wei lai zhuang tai de zhi biao fen zi ,jing zhun jian ce chu ji zai huan zhe ti nei de han liang ze you zhu yu yi sheng dui huan zhe jin hang zui you xiao de lin chuang zhen duan he jie ru zhi liao ,yin ci ,dui ai zheng xiang guan dan bai de ling min jian ce xian de you wei chong yao 。MUC1shi yi chong wei yu shang pi xi bao ding duan mo bu wei de tang dan bai ,hui zai yi sang shi ji xing de ai xi bao biao mian guo liang biao da ,yin ci ,ji zai ren ti nei de han liang ke yi zuo wei ai zheng zao ji zhen duan de zhi biao 。ran er zai xi bao ai bian de zao ji jie duan ,MUC1de han liang jiao di ,bu yi jian ce ,you dai tan suo jian bian 、gao xiao 、ling min de jian ce fang fa 。liang zi dian shi zui chang yong de ying guang tan zhen zhi yi ,kuan ji fa 、zhai fa she 、che cun ke diao yi ji gao liang zi chan lv de te dian shi de liang zi dian zai bu tong yan jiu ling yu de dao le an fan ying yong ,li ru :sheng wu cheng xiang 、guang xue bian ma 、sheng wu chuan gan deng 。kuo pei ti shi zhi dui mu biao fen zi you te yi xing qin he li de gua he gan suan huo zhe tai duan ,chang bei she ji wei jian ce tan zhen yong yu mu biao wu de te yi xing ling min jian ce 。ji yu kuo pei ti yu MUC1de te yi xing jie ge zuo yong ,ke jiang kuo pei ti she ji wei jian ce tan zhen yong yu MUC1de ling min jian ce 。ci xing wei qiu ye ke jian chen wei “ci zhu ”,ju you gao bi biao mian ji 、yi xiu shi 、yi fen li de te dian ,an fan de ying yong yu he suan 、dan bai zhi 、xiao fen zi wu zhi yi ji ai xi bao deng fang mian de jian ce 。za jiao lian shi fan ying (HCR)shi yi chong chang yong de xin hao fang da ji shu ,zai yin fa lian de zuo yong xia ,fa ka jie gou de DNAxu lie H1he H2ke zi zu zhuang za jiao cheng yi tiao chang de DNAshuang lian ,zai mo ren he he suan mei de can yu xia ,jin tong guo DNAji lian za jiao fan ying ji ke shi xian jian ce xin hao kuo zeng 。ben lun wen ji yu kuo pei ti dui dan bai te yi xing shi bie zuo yong bing jie ge za jiao lian shi fan ying (HCR)xin hao fang da ji shu ,ju li yong ci zhu yi fen li de te xing xian zhe jiang di bei jing gan rao ,she ji le liang chong liang zi dian ying guang tan zhen ,shi xian le dui MUC1de jian bian 、ling min ji te yi xing de jian ce ,zhu yao yan jiu nei rong ru xia :(1)can zhao ben ke ti zu yi you de fang fa ,zhi bei le CdZnTeSliang zi dian ;tong shi zai jian ce ti ji zhong yin ru he suan “fen zi guang kai guan ”liao (Ⅱ)pei ge wu —ji zai shui rong ye zhong ji ben mo ying guang xin hao ,dang qian ru DNAshuang lian zhong shi ,ke yi ce de ming xian de ying guang xin hao ;li yong liao (Ⅱ)pei ge wu dui liang zi dian de ying guang cu mie zuo yong ji liao (Ⅱ)pei ge wu qian ru DNAshuang lian shi ,dui liang zi dian mo cu mie zuo yong de te xing ,jiang ke te yi xing shi bie MUC1de kuo pei ti she ji wei fa ka jie gou ba xiang MUC1,qi dong za jiao lian shi fan ying (HCR)jiang xin hao fang da ,jing guo ci fen li hou ce ding shang qing ye de ying guang jiang du 。ju ci ,ke yi gen ju liao (Ⅱ)pei ge wu dui CdZnTeSliang zi dian de cu mie cheng du lai ding liang jian ce MUC1,xian xing fan wei 0.2-100 ng/mL(LOD=0.13 ng/mL,S/N=3),ci fang fa ye ke dui xie qing zhong MUC1han liang jin hang ling min ce ding 。(2)jiang qian yi zhang jian ce MUC1de fang fa jin yi bu gai jin ,mo xu yin ru liao (Ⅱ)pei ge wu 。cai yong yi guo fa zhi jie ge cheng DNAgong neng hua liang zi dian ,DNAgong neng hua liang zi dian tong guo yu hu bu chan lian za jiao yin fa za jiao lian shi fan ying xing cheng chang de shuang lian 。dang ti ji zhong cun zai MUC1shi ,ci qian bao chi wen ding fa ka jie gou de H0you yu yu MUC1de she die chan rao ,jie gou fa sheng gai bian ,bao lou chu chan lian mo duan ,bing ke yi yu fa ka jie gou de Fuelfa sheng hu bu za jiao ,jin er shi Fuelwei duan cheng wei zhi lian jie gou ,cong er ke yi yu da liang de S1he S2(DNA-CdTe:Zn2+QDs)fa sheng zhou ji de hu bu za jiao ,sheng cheng za jiao lian shi fan ying chan wu (shuang lian zhong han you da liang de liang zi dian )。jing guo ci fen li hou ,zhi jie jian ce shang qing ye de ying guang jiang du ,ji ke dui MUC1jin hang ding liang jian ce ,xian xing fan wei 0.15-100 ng/mL(LOD=0.11ng/mL,S/N=3),bing ke yong yu xie qing zhong MUC1de ling min jian ce 。
论文参考文献
论文详细介绍
论文作者分别是来自武汉大学的李政,发表于刊物武汉大学2019-07-22论文,是一篇关于量子点论文,杂交链式反应论文,磁珠论文,钌配合物论文,武汉大学2019-07-22论文的文章。本文可供学术参考使用,各位学者可以免费参考阅读下载,文章观点不代表本站观点,资料来自武汉大学2019-07-22论文网站,若本站收录的文献无意侵犯了您的著作版权,请联系我们删除。
标签:量子点论文; 杂交链式反应论文; 磁珠论文; 钌配合物论文; 武汉大学2019-07-22论文;