基于核酸连接等温扩增反应的癌症早期诊断新方法研究

项目名称： 基于核酸连接等温扩增反应的癌症早期诊断新方法研究

项目编号： No.21335005

项目类型： 重点项目

立项/批准年度： 2014

项目学科： 数理科学和化学

项目作者： 李正平

作者单位： 陕西师范大学

项目金额： 300万元

中文摘要： 癌症是一类与基因密切相关的重大疾病，其治疗的关键是实现早期诊断。利用基因分析进行癌症早期诊断的瓶颈是没有确证的基因标志物。根据与癌症发生、发展密切相关的多种因素，本项目提出以关键基因的突变、甲基化及相关的mRNA与microRNA作为综合的基因标志物研究癌症的早期诊断。同时本项目拟基于DNA探针连接形成的等温指数扩增反应，创新性地研究建立适合于基因突变、甲基化及mRNA、microRNA检测的分析技术平台。该分析技术仪器简单、操作方便，灵敏度达到能完全满足实际临床样品分析的水平；选择性达到有效检测单碱基差别核酸序列的水平。应用该项新技术，通过检测癌细胞与正常细胞中各种基因标志物的差别，确立适合癌症诊断的基因标志物；进一步通过实际临床诊断的检验，最后形成具有自主知识产权的癌症诊断试剂盒。

中文关键词： 等温扩增;基因诊断;癌症;DNA;RNA

英文摘要： The cancer is a class of heavy disease which is closely associated genes. The key problem for the cancer therapy is to realize the early diagnosis. However, the bottleneck of early diagnosis for cancer based on genetic analysis is lack of accurate genetic markers. Based on the multiplex factors closely related to cancer generation and development, we want to study the early diagnosis of cancers by using multiplex genetic markets, including the genetic mutation and methylation of the critical genes, and related messenger RNA, and microRNAs. Meanwhile, we will also study and establish a platform for analysis of genetic mutation, DNA methylation, messenger RNA, and microRNA by using ligation-based exponentially isothermal amplification of nucleic acids. With the analysis platform, the detection of multiplex genetic markers should be easily accomplished with simple instruments. The sensitivity should well meet the need for analysis of clinical samples, and the selectivity is enough to efficiently discriminate the single base difference in the sequences of nucleic acids. By using the analysis platform, we will determine the suitable genetic markers for early diagnosis of cancers through comparison of the different levels of various genetic markers between the cancer cells and the normal cells. The determined genetic marker will be further confirmed with the clinical diagnosis. Finally, we will develop the reagent kit under self-owned intellectual property for diagnosis of cancers.

英文关键词： Isothermal amplification;Genetic diagnosis;Cancer;DNA;RNA