本文選題：甲狀腺癌 + 基因�。� 參考：《吉林大學(xué)》2017年碩士論文

【摘要】：背景:甲狀腺癌在人類惡性腫瘤中約占1%,是最常見的內(nèi)分泌惡性腫瘤。在世界范圍內(nèi)發(fā)病率呈上升趨勢(shì),成為增長(zhǎng)最迅速的人類腫瘤。目前超聲、細(xì)針穿刺細(xì)胞學(xué)檢查(fine-needle aspiration cytology,FNAC)和血清甲狀腺球蛋白檢測(cè)等是甲狀腺腫瘤最常見的診斷方法,但因其存在著誤診和漏診的機(jī)率,建立更為準(zhǔn)確和特異的診斷方法成為臨床急需解決的問題。BRAF、RAS和TERT啟動(dòng)子區(qū)的突變因引起細(xì)胞異常增殖而在甲狀腺癌的發(fā)生中具有不可估量的作用,結(jié)合近年來分子診斷技術(shù)在腫瘤診治中的應(yīng)用,這些基因突變的檢測(cè)在甲狀腺癌中也已開展。但現(xiàn)有檢測(cè)技術(shù)存在著因醫(yī)師經(jīng)驗(yàn)不足和檢驗(yàn)設(shè)備落后等導(dǎo)致的假陽性和假陰性的情況,因此,一種新的高靈敏性和特異性的基因診斷方法有待研究。目的:建立新型甲狀腺癌相關(guān)基因檢測(cè)技術(shù),提高臨床檢測(cè)的準(zhǔn)確性,為臨床診治提供依據(jù)。方法:合成檸檬酸穩(wěn)定的金納米粒子用于構(gòu)建功能化金納米粒子的標(biāo)記探針;制備DNA微陣列芯片用于檢測(cè)靶標(biāo)DNA;通過銀增強(qiáng)反應(yīng)放大信號(hào);Array It Spot Ware Colorimetric微陣列掃描儀掃描DNA微陣列芯片獲得RLS信號(hào);Image J提取RLS信號(hào)值及背景信號(hào)值用以數(shù)據(jù)分析。結(jié)果:當(dāng)芯片點(diǎn)樣探針濃度分別為Probe 1T和1S:30μM,Probe2T、2S、3T、3S:5μM;雜交緩沖溶液為3×SSC,0.1%(w/v)SDS;功能化金納米粒子緩沖溶液為1×SSC,0.1%(w/v)SDS時(shí),45℃雜交反應(yīng)進(jìn)行為2小時(shí),30℃功能化金納米粒子反應(yīng)1小時(shí)的檢測(cè)條件下,可獲得較為穩(wěn)定和特異的檢測(cè)結(jié)果。結(jié)論:應(yīng)用DNA微陣列芯片結(jié)合功能化金納米粒子檢測(cè)甲狀腺癌相關(guān)基因是一種靈敏、特異的方法,對(duì)于臨床診治具有良好的參考價(jià)值。
[Abstract]:Background: thyroid carcinoma accounts for about 1% of human malignant tumors and is the most common endocrine malignant tumor. In the world, the incidence of disease is on the rise, becoming the fastest growing human tumor. At present, ultrasound, fine needle aspiration cytology (fine-needle aspiration) and serum thyroglobulin are the most common diagnostic methods for thyroid neoplasms. The establishment of more accurate and specific diagnostic methods has become an urgent clinical problem. Mutations in the BRAFRAS and TERT promoters play an inestimable role in the carcinogenesis of thyroid cancer due to abnormal cell proliferation. In combination with the recent application of molecular diagnostic techniques in the diagnosis and treatment of cancer, the detection of these gene mutations has been carried out in thyroid carcinoma. However, there are false positive and false negative due to the lack of doctors' experience and poor testing equipment. Therefore, a new method of gene diagnosis with high sensitivity and specificity needs to be studied. Objective: to establish a novel detection technique for thyroid oncogene, to improve the accuracy of clinical detection and to provide evidence for clinical diagnosis and treatment. Methods: citric acid stabilized gold nanoparticles were synthesized to construct labeled probes for functional gold nanoparticles. DNA microarray chip was prepared to detect target DNA, and RLS signal was obtained by scanning DNA microarray chip by array it spot microarray scanner. RLS signal and background signal were extracted for data analysis. Results: when the probe concentration was Probe1T and 1s: 30 渭 M, respectively, the hybridization reaction was carried out at 45 鈩，

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