發(fā)布時(shí)間：2018-06-16 22:27

  本文選題：表面增強(qiáng)拉曼散射 + 傳感器�。� 參考：《南京郵電大學(xué)》2017年碩士論文

【摘要】：核酸的表達(dá)失控與腫瘤的發(fā)生發(fā)展密切相關(guān),開(kāi)展腫瘤相關(guān)的核酸標(biāo)志物檢測(cè)有望實(shí)現(xiàn)腫瘤的早期檢測(cè)。然而在病變?cè)缙谀[瘤核酸標(biāo)志物含量極低,急需發(fā)展可靠的高靈敏檢測(cè)新方法。SERS技術(shù)具有超高靈敏度、非破壞性、抗光漂白,譜峰尖銳適合多組分檢測(cè)等優(yōu)異的性能使其成為分子檢測(cè)的重要手段。因此,發(fā)展并制備基于SERS的核酸標(biāo)志物傳感器及其多元聯(lián)合檢測(cè)技術(shù),對(duì)于實(shí)現(xiàn)腫瘤的早期檢測(cè)意義重大。針對(duì)這一需求,本論文基于SERS活性材料如銀納米棒陣列型基底、新型磁核枝杈狀金殼納米顆粒,設(shè)計(jì)并開(kāi)發(fā)了多種靈敏度、特異性俱佳的SERS傳感器,對(duì)血清中的痕量肺癌核酸標(biāo)志物進(jìn)行了快速、靈敏的聯(lián)合檢測(cè),為SERS技術(shù)在腫瘤早期檢測(cè)中的應(yīng)用并建立了技術(shù)平臺(tái),主要研究?jī)?nèi)容分為以下3個(gè)方面:(1)基于陣列型固態(tài)SERS基底和分子信標(biāo)發(fā)展了一種高靈敏多元mi RNA檢測(cè)傳感芯片。首先采用斜角沉積法蒸發(fā)鍍膜技術(shù)制備了大面積均一的高性能銀納米棒(Ag NR)陣列型SERS基底,并在基底表面修飾了三種標(biāo)記不同拉曼分子的DNA探針(分子信標(biāo)),構(gòu)建得到了mi RNA陣列型SERS傳感器,通過(guò)表征識(shí)別目標(biāo)mi RNA前后SERS強(qiáng)度的變化實(shí)現(xiàn)了對(duì)mi RNA的定性和定量檢測(cè)。該陣列型傳感器具有靈敏度和特異性高,均一性和穩(wěn)定性好,可重復(fù)使用等優(yōu)點(diǎn),可同時(shí)檢測(cè)血清中的三種肺癌標(biāo)志物mi RNA-21/486/375,檢測(cè)限分別達(dá)到393 a M、176 a M、144 a M。(2)基于磁核枝杈狀金殼納米顆粒結(jié)合四面體DNA探針,設(shè)計(jì)并構(gòu)建了mi RNA全液相SERS傳感器。首先基于種子生長(zhǎng)法并引入定向生長(zhǎng)控制劑Ag NO3,合成了一種磁核枝杈狀金殼納米顆粒;隨后設(shè)計(jì)組裝了四面體DNA探針,并將其修飾到磁核枝杈狀金殼納米顆粒表面構(gòu)建得到了兼具SERS活性和超順磁性的檢測(cè)基底;然后通過(guò)在金納米顆粒表面修飾單鏈DNA和拉曼活性分子DTNB構(gòu)建了SERS探針;最后利用堿基互補(bǔ)配對(duì)形成“檢測(cè)基底-mi RNA-SERS探針”三明治結(jié)構(gòu)復(fù)合物,借助外加磁場(chǎng)分離檢測(cè)液中的復(fù)合物并富集后進(jìn)行SERS檢測(cè),實(shí)現(xiàn)了對(duì)于血清中腫瘤核酸標(biāo)志物mi RNA-21的高靈敏、特異性檢測(cè),檢測(cè)限達(dá)到623 a M。(3)發(fā)展了基于四面體DNA核酸探針的多元SERS傳感器。針對(duì)性設(shè)計(jì)并組裝了具有特異性識(shí)別功能的四面體DNA核酸探針,并將四面體DNA核酸探針有序組裝到Ag NR陣列型基底表面構(gòu)建了高靈敏SERS核酸檢測(cè)基底;通過(guò)鏈置換反應(yīng),四面體DNA探針可同時(shí)識(shí)別兩種不同的目標(biāo)核酸,并捕獲兩種基于金納米球構(gòu)建的特異性SERS探針組裝成金納米球二聚體,形成SERS“熱點(diǎn)”;借助SERS探針信號(hào)強(qiáng)度實(shí)現(xiàn)兩種核酸分子的高靈敏檢測(cè)。該傳感器具有良好的特異性,線性檢測(cè)范圍寬(100 a M-1 n M),Target 1和2的檢測(cè)限分別為50 a M和66 a M。
[Abstract]:The uncontrolled expression of nucleic acid is closely related to the occurrence and development of tumor. The detection of tumor-related nucleic acid markers is expected to realize the early detection of tumor. However, the content of nucleic acid markers in tumor is very low in the early stage of pathological changes, so it is urgent to develop a new method of high sensitivity, non-destructive and anti-photobleaching. The sharp spectrum peak is suitable for multicomponent detection and so on, which makes it an important method for molecular detection. Therefore, the development and preparation of SERS-based nucleic acid marker sensors and multivariate combined detection techniques are of great significance for the early detection of tumors. In order to meet this demand, SERS active materials such as silver nanorod array substrate and new magnetic core branched gold shell nanoparticles are used to design and develop a variety of SERS sensors with high sensitivity and specificity. The rapid and sensitive combined detection of trace lung cancer nucleic acid markers in serum was carried out, which provided a platform for the application of SERS technique in early detection of tumor. Based on the array solid state SERS substrate and molecular beacons, a highly sensitive and multivariate sensor chip for mi RNA detection is developed. Firstly, a large area uniform silver nanorod array SERS substrate was prepared by the oblique angle deposition method. Three kinds of DNA probes (molecular beacons) labeled with different Raman molecules were modified on the substrate surface, and the mi RNA array SERS sensor was constructed. The qualitative and quantitative detection of mi RNA was achieved by characterizing the changes of SERS intensity before and after recognition of target mi RNA. The array sensor has the advantages of high sensitivity and specificity, good uniformity and stability, and can be reused. Three lung cancer markers, mi RNA-21 / 486 / 375, with a detection limit of 393 a / M ~ (176) a / M ~ (-1) ~ (144) a 路m ~ (2), respectively, were detected simultaneously. Based on the magnetic core branched gold shell nanoparticles and tetrahedral DNA probe, a full-liquid phase SERS sensor was designed and constructed. Based on the seed growth method and the introduction of Agno _ 3, a magnetic core branched gold shell nanoparticles were synthesized, and the tetrahedron DNA probe was designed and assembled. The SERS probe was constructed on the surface of the branched gold shell nanoparticles with both SERS activity and superparamagnetism, and the SERS probe was constructed by modifying single strand DNA and Raman active molecule DTNB on the surface of gold nanoparticles. Finally, the sandwich structure complex of "detecting substrate-mi RNA-SERS probe" was formed by complementary pair of bases, and the complex in the solution was separated and enriched by external magnetic field, and then SERS was detected. A multivariate SERS sensor based on tetrahedral DNA probe was developed for the detection of tumor nucleic acid marker mi RNA-21 with a detection limit of 623 a M. A tetrahedral DNA probe with specific recognition function was designed and assembled, and the tetrahedron DNA probe was sequentially assembled onto the surface of Ag NR array substrate to construct a highly sensitive SERS nucleic acid detection substrate. Tetrahedral DNA probes can simultaneously recognize two different target nucleic acids, and capture two specific SERS probes based on gold nanospheres to form gold nanospheres dimer to form "hot spots" of SERS. The signal intensity of SERS probe is used to detect the two kinds of nucleic acid molecules with high sensitivity. The sensor has good specificity and the detection limits of the linear detection range are 50 mm and 66 mm, respectively.
【學(xué)位授予單位】：南京郵電大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TP212

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 SHI Wen-sheng;;Surface-enhanced Raman scattering from semiconductor nanostructures[J];量子電子學(xué)報(bào);2014年01期

2 譚恩忠;殷鵬剛;郭林;;Fabrication of a network structure SERS substrate and the application in ultra-sensitive crystal violet detection[J];Optoelectronics Letters;2013年05期

3 顧仁敖;宗亞平;冷永章;凌德洪;;銀電極表面吸附異煙酸的表面增強(qiáng)喇曼散射(SERS)光的實(shí)驗(yàn)研究[J];光電子.激光;1988年06期

4 杜勇,方炎;鹵離子吸附競(jìng)爭(zhēng)對(duì)羅丹明B-金膠體系NIR-SERS的影響[J];光電子·激光;2003年08期

5 譚恩忠;;Gold nanoparticles-decorated single silver nanowire as an efficient SERS-active substrate[J];Optoelectronics Letters;2014年04期

6 凌德洪,周義新,過(guò)祥龍;吡啶在銀膠、氯化銀膠中的表面拉曼增強(qiáng)(SERS)[J];量子電子學(xué);1985年04期

7 龔繼來(lái);呂璞;曾光明;;表面增強(qiáng)拉曼光譜在環(huán)境分析中的研究進(jìn)展[J];化學(xué)傳感器;2009年03期

8 方靖淮;曹敏;成鳴飛;金永龍;;樹(shù)枝狀A(yù)g基底的制備及其SERS活性研究[J];光電子.激光;2010年11期

9 高美;方炎;;應(yīng)用置換反應(yīng)制備高效SERS活性基底研究[J];光電子.激光;2008年10期

10 蔡志鵬;王波;賀廷超;張玲;莫育俊;;對(duì)SERS襯底表面雜質(zhì)的處理研究[J];光散射學(xué)報(bào);2007年02期

相關(guān)會(huì)議論文 前10條

1 劉偉;秦維;劉可;袁亞仙;姚建林;顧仁敖;;一種新型可循環(huán)SERS基底的制備[A];中國(guó)化學(xué)會(huì)第27屆學(xué)術(shù)年會(huì)第04分會(huì)場(chǎng)摘要集[C];2010年

2 莫育俊;雷潔;李秀英;張鵬翔;;吸附于銅表面上分子的SERS及表面粗糙度對(duì)SERS的影響[A];全國(guó)第三屆光散射學(xué)術(shù)會(huì)議論文摘要[C];1985年

3 艾小紅;高小平;李秀英;莫育俊;;蘋果酸吸附在銅和銀表面上SERS的比較[A];全國(guó)第三屆光散射學(xué)術(shù)會(huì)議論文摘要[C];1985年

4 高敏俠;林秀梅;任斌;;均勻的強(qiáng)SERS活性基底的制備及表征[A];第十四屆全國(guó)分子光譜學(xué)術(shù)會(huì)議論文集[C];2006年

5 高美;方炎;;對(duì)樹(shù)枝狀銀納米結(jié)構(gòu)作為SERS增強(qiáng)基底的研究[A];第十四屆全國(guó)光散射學(xué)術(shù)會(huì)議論文摘要集[C];2007年

6 王春旭;陳雷;紀(jì)南;阮偉東;趙冰;趙純;;銀的無(wú)電沉積及其SERS活性的研究[A];第十五屆全國(guó)分子光譜學(xué)術(shù)報(bào)告會(huì)論文集[C];2008年

7 張鵬翔;付石友;龐玉璋;;表面增強(qiáng)喇曼散射的最近研究[A];中國(guó)物理學(xué)會(huì)光散射專業(yè)委員會(huì)成立十周年暨第六屆學(xué)術(shù)會(huì)議論文集（上）[C];1991年

8 ;SERS on thin film composed of gold nanoparticles[A];第十六屆全國(guó)光散射學(xué)術(shù)會(huì)議論文摘要集[C];2011年

9 楊U，

本文編號(hào)：2028318