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Detection and identification of emergence of HBV YMDD motif mutation during lamivudine treatment by hybridization to an oligonucleotide array
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摘要: 建立乙型肝炎病毒变异基因诊断芯片对拉米夫定治疗慢性乙型肝炎过程中出现的肝炎病毒P基因区YMDD变异进行快速准确的检测方法。设计特异性寡核苷酸探针数组 ,特殊处理芯片载体。用点样法制备乙型肝炎病毒变异基因诊断芯片。在本院住院治疗病人中选择 3 0例服用拉米夫定后 ,可能出现YMDD变异的病人进行基因芯片杂交检测分析 ;同时用PCR直接测序法对上述 3 0例病人血清标本进行双盲HBVDNA聚合酶活性区域测序对照。 3 0例服药后HBVDNA反跳病人中 ,基因芯片测得HBVYMDD变异 2 1例 ,其中YVDD变异 11例。YIDD变异 10例。HBVDNAPCR直接序列测定结果与基因芯片检测结果完全一致。乙型肝炎病毒变异基因诊断芯片可以同时检测YVDD、YIDD变异 ,同PCR直接测序法比较 ,准确率达 10 0 % ,无假阳性Abstract: 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%.
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Key words:
- HBV /
- HBV YMDD Motif Mutation /
- Lamivudine /
- Gene Chip
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