三代测序技术及其应用研究进展

Research Progress on Three Generations Sequencing Technology and Its Application

通过DNA测序不仅可以更好地认识生命的本质,了解生物的差异性、进化及发展史,而且为重组DNA的研究提供了方向,在疾病诊断及基因分型方面具有非常重要的实用价值。首先从发现DNA是遗传物质开始,简要回顾了第一、二和三代测序技术的整个发展历程,以及各自的优缺点和主要测序技术或平台;其次,重点介绍了由PacBio公司开发的第三代测序典型技术——单分子实时测序技术(SMRT)和ONT纳米孔测序技术的原理、方法和技术流程,并总结了第三代测序技术的应用领域,包括基因组重测序、do novo测序、转录组研究、甲基化检测、与疾病相关的结构变异检测,以及流行病学中病毒准种分析等应用;再次,比较了三代测序技术在转录组测序和表观遗传学研究中的技术优势,第三代测序技术在基因组重复区域或结构变异等研究领域具有非常明显的优势,对多种疾病的研究意义重大,已成为未来重要的精准诊断工具,此外,凭借对重要经济物种遗传信息的解析,育种工作者通过建立基因与性状的关联,对持有优良性状基因的个体进行人工选育,大大缩短了育种年限;最后,对第三代测序技术在医疗、农业、环境等领域的应用前景进行了展望。

DNA sequencing not only helps us better to understand the nature of life,biological differences,different biological evolutions and developmental history of organisms,but also provides direction for recombinant DNA research,and has very important practical value in disease diagnosis and genotyping.Firstly,from the discovery that DNA is a genetic material,we briefly review the entire development process of the first,second and third generation sequencing technologies,their respective advantages and disadvantages,and the main sequencing technologies or platforms in each.Secondly,we focus on the third generation sequencing typical technology developed by PacBio,including single molecule Real time(SMRT),Oxford Nanopore Technologies(ONT),introduce the principles,methods and technical processes,and summarize the application areas of the third generation sequencing technology,including the genomic resequencing,do novo sequencing,transcriptome studies,methylation testing,disease-related structural variation testing and the application of viral quasispecies analysis in epidemiology.Thirdly,the advantages of three generations of sequencing techniques in transcriptome sequencing and epigenetics are compared,the third generation sequencing technology has obvious advantages in research fields,such as genomic repeat regions or structural variations,and very meaningful for the study of various diseases,it is considered to be an important and accurate diagnostic tool in the future.In addition,breeders can artificially select individuals by establishing associations between genes and traits by analyzing the genetic information of important economic species,which greatly shortens the breeding period.Finally,we prospect the future application of the third generation sequencing technology in medicine,agriculture and environment.