|
|||||
|
|
| 锁核酸探针多重荧光定量PCR检测HBV基因变异 | |
| 引用本文： | 江洁,丁渭,陈华云,何琼,何蕴韶. 锁核酸探针多重荧光定量PCR检测HBV基因变异[J]. 广东医学, 2011, 32(1) |
| 作者姓名： | 江洁 丁渭 陈华云 何琼 何蕴韶 |
| 作者单位： | 中山大学医学院基础医学院临床检验诊断专业 中山大学达安基因诊断中心 |
| 基金项目： | 国家科技部863计划项目 |
| 摘 要： | [摘要] 目的 描述并评价锁核酸（locked nucleic acid,LNA）探针实时荧光定量聚合酶链反应（Real-Time PCR）检测乙型肝炎病毒（HBV）204位基因突变的方法及特点。方法 分别用直接测序法、锁核酸（LNA）探针实时荧光定量PCR检测109例接受拉米夫定（LAM）治疗6个月以上的慢性乙型肝炎患者的血清样本。结果 LNA探针实时荧光定量PCR能检测HBV野生型YMDD和其突变型YVDD,YIDD。而且，这种方法还可以检测HBV野生型和变异型的混合。LNA探针相比普通探针在检测野生型和突变型混合样本方面，更敏感，区分度更好。结论 LNA探针实时荧光定量PCR检测YMDD变异具有快速，灵敏，等特点，可以用于大量临床样本的快速筛查，在监测拉米夫定治疗疗效及发现新耐药突变型等方面具有广泛的应用前景。 |
| 关 键 词： | 锁核酸探针 多重荧光定量PCR YMDD 乙型肝炎病毒 拉米夫定耐药突变 |
Multiple locked nucleic acid probes to detect hepatitis B virus mutations using real-time PCR |
|
| Abstract: | 【Objective】To evaluate and describe the advantages of real-time polymerase chain reaction(PCR) using locked nucleic acid(LNA) probes in detecting the mutations of hepatitis B virus. 【Methods】One hundred and nine serum samples used in this research were obtained from 109 patients with chronic hepatitis B virus infection, who had received Lamivudine therapy for at least six months. All specimens were detected by direct sequencing and multiplex locked nucleic acid probes real-time PCR. 【Results】LNA probe real-time PCR can detect wild type tyrosine-methionine-aspartate-aspartate(YMDD),and its mutants, tyrosine-isoleucine-aspartate-aspartate(YIDD) and tyrosine-valine-aspartate-aspartate(YVDD).Besides, the mixtures of the wild-type virus and the mutants in the serum sample could be detected. Compared to normal probes, the LNA probes real-time PCR is more sensitive in detection of mixed samples. 【Conclusion】The real-time PCR with LNA probes is a less time-consuming, a more sensitive and specific method to detect the mutations at the YMDD motif, and can be widely used in a large number of the clinical samples for fast-detecting. It’s also a perspective method for monitoring lamivudine effect and a novel viral mutants in patients undergoing lamivudine antiviral therapy. |
| Keywords: | YMDD |
| 本文献已被 万方数据 等数据库收录！ | |
| 点击此处可从《广东医学》浏览原始摘要信息 | |
| 点击此处可从《广东医学》下载全文 | |