本文選題：單核苷酸多態(tài)性 + 生物傳感器　； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：單核苷酸多態(tài)性(single nucleotide polymorphisms,SNP)是被公認(rèn)的導(dǎo)致基因多樣性與復(fù)雜性的重要原因。β1腎上腺素能受體基因(ADRB1)發(fā)生SNP直接影響高血壓患者服用美托洛爾的降壓效應(yīng)。而維生素K環(huán)氧化物還原酶復(fù)合物亞基1(VKORC1)基因發(fā)生SNP與華法林的使用劑量有密切關(guān)系。因此,建立全新的SNP分子檢測(cè)方法可以更好地掌握針對(duì)某種藥物反應(yīng)性的個(gè)體間差異,從而實(shí)現(xiàn)個(gè)體化治療,降低醫(yī)療風(fēng)險(xiǎn)和醫(yī)療費(fèi)用。本文通過合成全新的納米材料,用于修飾電化學(xué)DNA傳感器,實(shí)現(xiàn)了對(duì)兩種比較常見的臨床應(yīng)用藥物相關(guān)的SNP的檢測(cè),以期精準(zhǔn)指導(dǎo)個(gè)體化臨床用藥。研究工作分為以下兩部分:第一章應(yīng)用氧化石墨烯/二氧化鈰/鉑納米粒子作為納米信標(biāo)對(duì)β1腎上腺素能受體基因的電化學(xué)檢測(cè)本實(shí)驗(yàn)構(gòu)建了一個(gè)基于氧化石墨烯/二氧化鈰/鉑納米粒子(GO/CeO2/PtNPs)作為納米信標(biāo),金納米粒子/鏈霉親和素(Au NPs/SA)作為電極修飾納米材料形成的DNA傳感器,用于血清中ADRB1突變特異性檢測(cè)。首先以GO為載體,固載了大量的CeO2。接著,在其表面還原了Pt NPs,合成了GO/CeO2/PtNPs獲得良好的催化性能。另外,在玻碳電極(GCE)表面沉積一層AuNPs之后修飾SA來連接生物素標(biāo)記的探針。通過各種條件優(yōu)化,制備的傳感器靈敏度較高,特異性較好,線性范圍較寬,重現(xiàn)性較好。線性范圍為分為兩段:1 fM到100 fM和100 fM到10 nM,最低檢測(cè)限為0.33 fM。此外,加樣回收率在99.4-101.9%之間,說明該傳感器可以應(yīng)用于血清中ADRB1基因突變的檢測(cè)。第二章應(yīng)用富勒烯-聚酰胺-胺型樹枝狀高分子-空心鉑作為納米信標(biāo)對(duì)維生素K環(huán)氧化物還原酶復(fù)合物亞基1的電化學(xué)檢測(cè)本實(shí)驗(yàn)構(gòu)建了一個(gè)基于富勒烯-聚酰胺-胺型樹枝狀高分子-空心鉑(C60NPs-PAMAM-PtPNPs)作為納米信標(biāo),還原性氧化石墨烯四乙烯五胺/金納米顆粒/鏈霉親和素(rGO-TEPA/AuNPs/SA)作為電極修飾納米材料形成的DNA傳感器,用于血清中VKORC1基因突變的特異性檢測(cè)。C60NPs不僅具有氧化還原活性可以作為信號(hào)物質(zhì),而且還能作為生物分子的載體來固載PtPNPs。因此,我們合成了C60NPs-PAMAM-PtPNPs。在GCE表面修飾rGO-TEPA后再沉積一層AuNPs,接著修飾SA來連接生物素標(biāo)記的四面體DNA用于捕獲目標(biāo)DNA。通過條件優(yōu)化,傳感器的靈敏度較高,特異性較好,線性范圍較寬,重現(xiàn)性較好。線性范圍為1 pM到10 nM,最低檢測(cè)限為33 fM。此外,加樣回收率在96.7-103.94%之間,說明該傳感器可以應(yīng)用于血清中VKORC1基因突變的檢測(cè)。
[Abstract]:Single nucleotide polymorphisms (SNPs) are recognized as an important cause of gene diversity and complexity. 尾 1 adrenoceptor gene (ADRB1) has a direct impact on the antihypertensive effect of metoprolol on hypertension patients with SNP. The SNP of vitamin K epoxide reductase complex subunit 1 (VKORC1) was closely related to the dosage of warfarin. Therefore, the establishment of a new SNP molecular detection method can better grasp the individual differences in response to a certain drug, so as to achieve individualized treatment, reduce medical risks and medical costs. In this paper, a novel nano-material was synthesized to modify the electrochemical DNA sensor, and the detection of two common clinical drug related SNP was realized, in order to guide the individualized clinical medicine accurately. The research work is divided into two parts as follows: in Chapter 1, a novel method for the electrochemical detection of 尾 _ 1-adrenergic receptor gene by using graphene oxide / cerium oxide / platinum nanoparticles as a nano-beacon has been developed. A novel method based on graphite oxide has been developed for the electrochemical detection of 尾 _ 1-adrenergic receptor gene. The nanocrystalline CEO _ 2 / PtNPs / cerium dioxide / platinum nanoparticles are used as nanoscale beacons. Gold nanoparticles / streptavidin / au NPs / SAs were used as DNA sensors modified with electrodes to detect ADRB1 mutation specificity in serum. At first, a large amount of CEO _ 2 was immobilized on go carrier. Then, Pt NPs were reduced on the surface and GO/CeO2/PtNPs was synthesized to obtain good catalytic performance. In addition, a layer of AuNPs was deposited on the surface of the glassy carbon electrode (GCE) and modified with SA to bind the biotin labeled probe. By optimizing various conditions, the sensor has high sensitivity, good specificity, wide linear range and good reproducibility. The linear range is divided into two segments: 1: 1 FM to 100 FM and 100 FM to 10 nm. The minimum detection limit is 0.33 fM. In addition, the recovery rate was 99.4-101.9%, which indicated that the sensor could be used to detect the mutation of ADRB1 gene in serum. Chapter 2 electrochemical detection of vitamin K epoxide reductase complex subunit 1 using fullerene-polyamide-amine dendrimer and hollow platinum as nanometer beacon Amino-amine type dendritic macromolecule, C60NPs-PAMAM-PtPNPs), used as nanoscale beacons, Reductive graphene tetraethylenetetramethylamine / gold nanoparticles / streptavidin / rGO-TEPA / AuNPs / SA) were used as DNA sensors for the modification of nanomaterials. The specific detection of VKORC1 gene mutation in serum. C60 NPs not only have redox activity but also can be used as carriers of biomolecules to immobilize PtPNPs. Therefore, we synthesized C60NPs-PAMAM-PtPNPs. A layer of AuNPs was deposited after rGO-TEPA was modified on the surface of GCE, and then SA was modified to bind biotin labeled tetrahedron DNA to capture target DNAs. The sensor has high sensitivity, good specificity, wide linear range and good reproducibility. The linear range is from 1 pm to 10 nm, and the minimum detection limit is 33 FM. In addition, the recovery rate was 96.7-103.94%, which indicated that the sensor could be used to detect the mutation of VKORC1 gene in serum.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R96;TP212.3

【參考文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

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本文編號(hào)：1880262