摘要
为探索一种快速、简便检测乙型肝炎病毒 (HBV)前 C区基因变异的方法 ,用聚合酶链反应 (PCR)技术扩增 94例乙肝患者血清 HBV前 C区基因片断 ,阳性扩增产物用单链构象多态性 (SSCP)法进行检测 ,对具有不同特征性电泳图谱的 PCR产物再以直接测序法加以核定 ,以辨别不同图谱与野生株、变异株或野生株、变异株混合感染之间的关系。结果显示 :有 86例患者 HBV前 C区 PCR扩增阳性 ,SSCP法检测发现有 3种常见的特征性电泳图谱 ,测序后证实 ,它们分别代表了野生株、变异株 (nt1896位 G→ A)及野生株、变异株混合感染的 SSCP图谱 ;在某些变异株图谱中 ,尚可见其它位点也存在变异 ,其中以 nt1899位 G→ A突变最为常见。提示 :以 PCR- SSCP技术检测 HBV前C区变异具有敏感性高 ,特异性强 ,简单、快捷、价廉等优点 ,而且特别适用于大样本的筛检 ,因此有应用价值 。
In order to find a more beneficial method for detecting the mutation of HBV precore gene, specific primers were designed to amplify HBV precore fragment in 94 serum samples of patients with hepatitis B. Subsequently, all positive PCR products were analyzed by single strand conformational polymorphism (SSCP) technique. According to the patterns of single strand DNA bands yielded in gels, sequential sequencing was performed on those PCR products with characteristic patterns of single strand bands. Precore fragments of HBV could be detected in 86 serum samples by PCR. The analysis of SSCP showed three mainly different patterns of single strand DNA bands in gels. Sequencing confirmed that they indicated wild type strain, mutant strain (at nt 1896 G→A) and mixed strains, respectively. In some mutant strain SSCP patterns, besides the mutation at nt 1896G→A, mixed mutants could also be observed commonly, for example at nt 1899 G→A. The results suggest both sensitivity and specificity are satisfactory for detecting HBV precore mutation by PCR SSCP technique. Furthermore, it possesses a few more beneficial advantages, such as quickness, simplicity, cheapness and fitness especially when used to detect a number of samples. So it may be recommended as a valuable technique to detect the mutation of HBV precore gene.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2002年第4期393-396,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
关键词
聚合酶链反应
单链构象多态性
乙型肝炎病毒
变异
polymerase chain reaction
single strand conformational polymorphism
hepatitis B virus
mutation
