[Analysis of three lamivudine-resistant HBV mutants with the method of restriction enzyme digestion and its application].
OBJECTIVE
To investigate the prevalence of three lamivudine-resistant HBV mutants in lamivudine-treated Chinese patients.


METHODS
Using three pairs of of HBV polymerase gene B and C domain fragments were amplified by PCR. The PCR products were then digested by restriction enzyme Nde I and Nla III. The digested products were analyzed by electrophoresis. With this method, the prevalence of the three lamivudine-resistant mutants in lamivudine-treated Chinese patients was investigated.


RESULTS
After Nde I digestion of p24 and p29 amplified product, HBV wild type could be easily separated from YMDD mutant. At the same time, YIDD could be separated from YVDD mutant after Nla III digestion of p24 and p29 amplified product. By this method, the authors found that these eleven patients were infected with lamivudine-resistant mutants. Six of them were infected with M5501 mutant; five were infected with M550V mutant (one of them had both M550V and L526M mutations).


CONCLUSIONS
The method of the present study was demonstrated to be an easy way to detect HBV lamivudine-resistant mutants and can be applied to clinical monitoring of lamivudine resistance.
Sun
Jian
J
Department of Infectious Diseases, Nanfang Hospital, Guangzhou 510515, China.
Hou
Jin-lin
JL
Xiao
Lei
L
Wang
Zhan-hui
ZH
Zhang
Lian
L
Luo
Kang-xian
KX
D000293:Adolescent; D000328:Adult; D000998:Antiviral Agents / Q000494:pharmacology / Q000627:therapeutic use*; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis; D024882:Drug Resistance, Viral / Q000235:genetics*; D005260:Female; D006509:Hepatitis B / Q000188:drug therapy / Q000821:virology; D006515:Hepatitis B virus / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology / Q000627:therapeutic use*; D008297:Male; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D017354:Point Mutation; D016133:Polymerase Chain Reaction
D000998:Antiviral Agents; D017931:DNA Primers; D004279:DNA, Viral; D019259:Lamivudine