title: [Detection of the mutation in HBV polymerase gene by RFLP PCR method in hepatitis B patients treated with lamivudine].
abstract: BACKGROUND
  To investigate the mutation of HBV polymerase gene in chronic hepatitis B patients treated with lamivudine.
  
  
  METHODS
  The restriction-fragment-length-polymorphism (RFLP) assay for HBV DNA sequence determination at the codon 528 and 552 in the HBV polymerase gene associated with lamivudine resistance in vitro. HBV DNA samples extracted from sera of 240 patients were subjected to PCR amplification with primer pairs F2/R2 (552), F3/R2 (528). Each PCR product was digested with Nde I or Nla III.
  
  
  RESULTS
  Serum HBV DNA mutation was found in 51/240 patients (38/51M552V, 26/38L528M, 13/51M552I) after therapy for 52 weeks. DNA sequence analysis was performed on samples of 3 patients, and the results were consistent with those of RFLP assay.
  
  
  CONCLUSIONS
  The RFLP assay was able to detect the mutation of HBV DNA at codon 552 and 528 which are the principal site of HBV DNA resistant to lamivudine. The specific PCR method for HBV DNA mutation is rapid, simple and specific.
authors.affiliation: Beijing Youan Hospital, Beijing 100054, China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Li
authors.forename: Yan-bin
authors.initials: YB
authors.lastname: Guo
authors.forename: Wa
authors.initials: W
authors.lastname: Hao
authors.forename: Zun-hui
authors.initials: ZH
authors.lastname: Lin
authors.forename: Hai-ying
authors.initials: HY
authors.lastname: Jin
authors.forename: De-gong
authors.initials: DG
authors.lastname: Liu
mesh_terms: D024882:Drug Resistance, Viral; D015684:Gene Products, pol / Q000235:genetics*; D006515:Hepatitis B virus / Q000201:enzymology / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000188:drug therapy* / Q000821:virology; D006801:Humans; D019259:Lamivudine / Q000627:therapeutic use*; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D018894:Reverse Transcriptase Inhibitors / Q000627:therapeutic use*
chemical_list: D015684:Gene Products, pol; C066027:P protein, Hepatitis B virus; D018894:Reverse Transcriptase Inhibitors; D019259:Lamivudine