分化型甲状腺癌相关基因微阵列基因芯片检测方法的构建

发布时间：2018-07-04 14:26

  本文选题：甲状腺癌 + 基因　； 参考：《吉林大学》2017年硕士论文

【摘要】：背景:甲状腺癌在人类恶性肿瘤中约占1%,是最常见的内分泌恶性肿瘤。在世界范围内发病率呈上升趋势,成为增长最迅速的人类肿瘤。目前超声、细针穿刺细胞学检查(fine-needle aspiration cytology,FNAC)和血清甲状腺球蛋白检测等是甲状腺肿瘤最常见的诊断方法,但因其存在着误诊和漏诊的机率,建立更为准确和特异的诊断方法成为临床急需解决的问题。BRAF、RAS和TERT启动子区的突变因引起细胞异常增殖而在甲状腺癌的发生中具有不可估量的作用,结合近年来分子诊断技术在肿瘤诊治中的应用,这些基因突变的检测在甲状腺癌中也已开展。但现有检测技术存在着因医师经验不足和检验设备落后等导致的假阳性和假阴性的情况,因此,一种新的高灵敏性和特异性的基因诊断方法有待研究。目的:建立新型甲状腺癌相关基因检测技术,提高临床检测的准确性,为临床诊治提供依据。方法:合成柠檬酸稳定的金纳米粒子用于构建功能化金纳米粒子的标记探针;制备DNA微阵列芯片用于检测靶标DNA;通过银增强反应放大信号;Array It Spot Ware Colorimetric微阵列扫描仪扫描DNA微阵列芯片获得RLS信号;Image J提取RLS信号值及背景信号值用以数据分析。结果:当芯片点样探针浓度分别为Probe 1T和1S:30μM,Probe2T、2S、3T、3S:5μM;杂交缓冲溶液为3×SSC,0.1%(w/v)SDS;功能化金纳米粒子缓冲溶液为1×SSC,0.1%(w/v)SDS时,45℃杂交反应进行为2小时,30℃功能化金纳米粒子反应1小时的检测条件下,可获得较为稳定和特异的检测结果。结论:应用DNA微阵列芯片结合功能化金纳米粒子检测甲状腺癌相关基因是一种灵敏、特异的方法,对于临床诊治具有良好的参考价值。
[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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