Tumor development is usually related to the genetic mutation and abnormal expression of multiple genes. Comprehensive analysis of tumor genome, transcriptome and epigenetics is very important for the rapid identification of disease-specific gene clusters and modification sites. Previously, the next-generation sequencing technology was mainly used to explore the information of genomes, however, it cannot meet the requirement of mechanical researches due to several problems such as difficult sequence assembly and leak detection of the low abundance factors. Therefore, single molecule sequencing technology gradually emerged with its unique superiority. This paper reviews the research processes of single molecule sequencing technology in several human tumors, and prospecs its application in clinical diagnosis.