[Detection of the mutation in HBV polymerase gene by RFLP PCR method in hepatitis B patients treated with lamivudine].
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.
Li
Zhuo
Z
Beijing Youan Hospital, Beijing 100054, China.
Guo
Yan-bin
YB
Hao
Wa
W
Lin
Zun-hui
ZH
Jin
Hai-ying
HY
Liu
De-gong
DG
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*
D015684:Gene Products, pol; C066027:P protein, Hepatitis B virus; D018894:Reverse Transcriptase Inhibitors; D019259:Lamivudine