拉米夫定治疗慢性乙型肝炎过程中HBV YMDD变异及基因芯片检测

To set up using the gene chip technology to detect and identify quickly and accurately the HBV P gene YMDD Motif mutation during the chronic hepatitis treated with lamivudine.DNA microarrays were prepared by spotting fluorescence labeled probes of target genes onto specially lattice glass slides with robotics.The serum samples of 30 patients with hepatitis B after 68 weeks treated with lamivudine were detected double-blind by gene chips and by nucleotide sequences assay technique to identify the rate of emergence of HBV P gene YMDD Motif Mutation.By the gene chips there were 21 patients with HBV P gene YMDD Motif Mutation including 11 cases with YVDD and 10 cases with YIDD Motif Mutation.By directly sequencing of PCR products there were 11 cases of YVDD with adenine 741 changed into cytidine resulted in methionine 552 changed into valine in which 6 cases with adenine 669 changed into cytidine and leucine changed into methionine 10 cases of YIDD Motif Mutation with guanosine 743 altered thymidine methionine 552 changed into isoleucine including 3 cases with thymidine 281 changed into cytidine and leucine 565 altered proline.The gene chip can be used to test HBV YVDD?YIDD Motif Mutation comparing with nucleotide sequences assay technique, the accuracy rate was 100%.