Objective To develop a rapid specific method to detect HBV gene mutation by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Methords The patients of YMDD and YIDD mutation were detected by DNA sequencing.Results the DNA sequencing demonstrated that not only YMDD and YIDD/YVDD mutation, but LLLL,LMLL,DLHD,DMHD,LLAQ and LMAQ mutation were also detected in different hepatitis B patients. The results of PCR-RFLP indicated that there were different restricted sites in the distinct mutation.Conclusion The results above indicated that all these mutants exist in the initiator codon of ATG(Met). The mutation of LMLL DMHD LMAQ was associated with Lamivordin treatment. Whether mutation of Met links with the second replicating of HBV needs to be further studied.