反向斑点杂交技术检测HBV YMDD基序变异的应用和评价  

Application and Evaluation of Reverse Dot Blot (RDB) Hybridization Assay for HBV YMDD Motif Mutants

作　　者：张太松[1] 董瑞华[1] 李建芳[1] 林炳生[1] 雍万军[1] 胡守旺[1] 李明 周新宇[1] 

机构地区：[1]中山大学达安基因诊断中心∥卫生部医药生物工程技术研究中心,广东广州510665

出　　处：《中山大学学报（医学科学版）》2009年第4期428-432,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广州市科技局科技攻关重大项目(2007Z1-E4031)

摘　　要：【目的】采用反向斑点杂交技术对拉米夫定治疗后的慢性乙型肝炎患者血清中HBV YMDD基序变异进行检测,并对其临床应用进行方法学评价。【方法】提取242例慢性乙型肝炎患者血清中HBV DNA,经聚合酶链反应后进行膜条杂交并同测序比较分析检测结果的一致性,随后对该方法进行灵敏度和混合感染检测能力的评价。【结果】242例样本检测结果有236例与测序结果相符,反向斑点杂交检测结果同测序结果的符合率达97.5%;混合型感染58例,占样品总数的24%。反向斑点杂交技术检测HBV YMDD基序变异的灵敏度达103IU/mL,在病毒混合感染群体中能检测出约占10%的突变型病毒株。【结论】反向斑点杂交技术检测HBV YMDD基序变异具有简单、准确、经济实用的特点,具有很好的临床应用前景。[Objective ] To detect HBV YMDD motif mutants using RDB hybridization assay in lamivudine treated patients with chronic hepatitis B virus infection, as well as to evaluate the detection capability for clinical application. [ Method] HBV DNA was extracted from serum for a total of 242 cases, after the PCR amplification, the hybridization was performed. By comparing the RDB assay results to sequence analysis, the concordant results were analyzed. The sensitivity and detection capability for mixed infection samples are also evaluated. [Results] There are 236 of concordant results for RDB assay and sequencing were obtained in a total of 242 cases, accounting for 97.5%. For all of the cases, there are 58 cases with coexisting mutant viruses in wild type viruses, accounting for 24%. The sensitivity of RDB hybridization assay for HBV YMDD motif mutants was 103 IU/mL, and approximately 10% mutant type strains can be detected from a mixed infection sample. [Conclusion] The RDB hybridization assay for HBV YMDD motif mutants is a simple, accurate, and economic method and it may be a promising tool for clinical application.

关 键 词：乙型肝炎病毒(HBV) 突变 耐药 反向斑点杂交 

分 类 号：R734.5[医药卫生—肿瘤]