随着基因测序技术的创新和应用，新的高通量测序技术不断涌现，以Pacific Biosciences(PacBio)公司的单分子实时测序(single molecule real time sequencing)为代表的第三代测序(third generation sequencing，TGS)技术开始逐渐应用于基因组研究，包括大型基因组拼装、基因结构变异和表观遗传研究等方面。本文主要对TGS技术的原理、特点和应用，特别是在病毒研究中的应用进行介绍，并与第二代测序(next generation sequencing，NGS)技术进行比较，为基因组测序技术的选择及其临床应用提供一定参考。

With the consecutive innovation and application of gene sequencing technology, the third generation sequencing (TGS) based on single molecule real time (SMRT) sequencing technology has demonstrated a potential in a variety of applications in basic and clinical studies, including whole genome assembly, detection of gene structure variation, epigenetic research, etc. This review mainly introduces the principle, characteristics and applications of the TGS technology, especially its applications in virus researches. This review also compares the differences between the next generation sequencing (NGS) and TGS, so as to provide a reference for application based selection for sequencing technology.