Li Zhuo, Guo Yan-bin, Hao Wa, Lin Zun-hui, Jin Hai-ying, Liu De-gong
Beijing Youan Hospital, Beijing 100054, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2003 Sep;17(3):266-9.
To investigate the mutation of HBV polymerase gene in chronic hepatitis B patients treated with lamivudine.
The restriction-fragment-length-polymorphism (RFLP) assay for HBV DNA sequence determination at the codon 528 and 552 in the HBV polymerase gene associated with lamivudine resistance in vitro. HBV DNA samples extracted from sera of 240 patients were subjected to PCR amplification with primer pairs F2/R2 (552), F3/R2 (528). Each PCR product was digested with Nde I or Nla III.
Serum HBV DNA mutation was found in 51/240 patients (38/51M552V, 26/38L528M, 13/51M552I) after therapy for 52 weeks. DNA sequence analysis was performed on samples of 3 patients, and the results were consistent with those of RFLP assay.
The RFLP assay was able to detect the mutation of HBV DNA at codon 552 and 528 which are the principal site of HBV DNA resistant to lamivudine. The specific PCR method for HBV DNA mutation is rapid, simple and specific.
研究拉米夫定治疗的慢性乙型肝炎患者中乙肝病毒聚合酶基因的突变情况。
采用限制性片段长度多态性（RFLP）分析法，对体外与拉米夫定耐药相关的乙肝病毒聚合酶基因第528和552密码子处的乙肝病毒DNA序列进行测定。从240例患者血清中提取的乙肝病毒DNA样本，用引物对F2/R2（552）、F3/R2（528）进行PCR扩增。每个PCR产物用Nde I或Nla III酶切。
治疗52周后，240例患者中有51例（38/51为M552V，26/38为L528M，13/51为M552I）血清乙肝病毒DNA发生突变。对3例患者的样本进行了DNA序列分析，结果与RFLP分析结果一致。
RFLP分析法能够检测到乙肝病毒DNA第552和528密码子处的突变，这两个位点是乙肝病毒对拉米夫定耐药的主要位点。用于检测乙肝病毒DNA突变的特异性PCR方法快速、简便且具有特异性。
