随着遗传病和表型性状相关的更多基因位点逐步被发现，诸多突变热点的快速高通量分析对临床诊断、风险评估、个体化治疗等尤为重要。本项目拟以地贫基因为研究靶点，以MALDI-TOF质谱为技术平台，采用目标基因模板多重PCR富集结合位点特异性质谱探针单碱基延伸的检测策略，建立同一反应体系中全突变热点基因分型新方法。此方法快速准确、高通量低成本，且操作简单能实现自动化和标准化，以满足我国南方地区地贫基础研究及人群防控的检测方法需求。同时从理论上建立MALDI-TOF质谱DNA分析的多基因、多位点、多突变类型快速基因分型技术方案和检测策略，从实践上取得解决高GC复杂二级结构及高同源序列影响的有效措施，为以地贫为代表的遗传病分子诊断方法学研究提供指导和借鉴，也可促进DNA质谱平台在遗传检测领域的推广应用。

As genetic diseases and phenotype related loci gradually discovered, rapid high-throughput analysis of numerous hotspot mutations is particularly important for clinical diagnosis, risk assessment, and individualized treatment. In this project, the thalassemia served as research target and the MALDI-TOF mass spectrometry as technique platform, a new method for whole thalassemia hotspot mutations genotyping in same reaction system should be developed，which based on the strategy of target gene template multiplex PCRs enrich combined with site-specific mass spectrometry probes single base extension. The method is rapid, accurate, high-throughput, low-cost, simple operation and can realize automation and standardization, which meets the detection method requirement of basic research and prevention programs of thalassemia in southern China. Meanwhile, a technical scheme and detection strategy for rapid genotyping of multi-genes, multi-sites, multi-mutation types with MALDI-TOF mass spectrometry DNA analysis platform should be theoretically established. Effective measures to settle the effects of high GC complex secondary structures and high sequence homologous, can provide reference for the molecular diagnostic methodological research of genetic diseases represented by thalassaemia, and also promotes the application of DNA mass spectrometry platform in the genetic testing field.