Abstract: The high mutation rate during hepatitis B virus( HBV) replication leads to HBV quasispecies. The study of HBV quasispecies has an important significance for the hepatitis B pathogenesis,prognosis,and outcome prediction. Recently,the next- generation sequencing( NGS) is extensively used in many biological and medical fields due to its high throughput,ultra- deep coverage,and high sensitivity,which also brings new strategies to HBV quasispecies studies. This article describes the working principles and features of several commonly used NGS technologies,reviews the application of NGS technologies in HBV quasispecies detection in recent years,and particularly discusses the variations in different regions of HBV genome and the population characteristics of HBV quasispecies. For now,NGS technologies used in HBV variation detection mainly presents the advantages of high sensitivity and high throughput. However,how to lower the cost,increase the accuracy of sequencing,and standardize the procedure of massive sequencing data management and personalized analysis are still challenging problems. The issue about NGS that matters at present and urgently needs to be solved is how to overcome its limitations,and then put it into HBV- related studies and ultimately clinical application.