摘要
目的 分析乙型肝炎病毒 (HBV)C基因启动子 (BCP)双突变与乙型肝炎临床表现及HBeAg表型的关系。方法 针对BCP双突变的特点 ,设计一条通用捕捉探针、一条野生型和一条双突变显色探针。待测标本DNA经聚合酶链反应 (PCR)扩增后分别与野生型和双突变显色探针杂交 ,然后用酶联免疫吸附试验 (ELISA)显示杂交结果 ,判定BCP突变与否。结果 14 7例经证实为HBVDNA阳性的急慢性肝炎患者中共有 5 1例双突变 ,其中 4 2例为单纯双突变 ,9例为混合突变 (双突变和野生型皆为阳性 )。 117例慢性中、重型肝炎中共有 36例BCP双突变 ,8例混合突变。 78例HBeAg阳性患者中 ,有 2 5例BCP双突变 ,其中 7例为混合突变。 6 5例HBeAg阴性患者中 ,有 2 6例BCP双突变。结论 慢性肝炎BCP双突变高于急性肝炎 ;BCP双突变对HBeAg的表达有一定影响 ;PCR微板核酸杂交ELISA技术是一种简便。
Objective To investigate 1762T、1764A double position mutation in the Basic Core Promoter(BCP) of hepatitis B virus and reveal its relation to clinical symptoms and HBeAg phenotype. Methods microplate sandwich hybridization technique was used to detect BCP double position mutation. One common capture probe and one mutant specific detector probe as well as one wild type detector probe were synthesized and hybridized with amplified HBV DNA from the sera of hepatitis B patients. The results of hybridization were exhibited with ELISA. Results 147 hepatitis B patients who had been confirmed HBV DNA positive were screened. 51 patients were BCP double position mutation, 42 of which were BCP single position mutation and, 9 were mix mutation(both mutation and wild type were positive). BCP mutant was detected in 36 of 117 with chronic hepatitis and, 8 of which were mix mutants. Moreover, BCP mutant was detected in 7 of 30 with acute hepatitis in 25 of 78 with HBeAg positive were mutant and in 26 of 65 with HBeAg negative were mutant.Conclusions (1) The rate of BCP double position mutation in chronic hepatitis B patients is higher than that in acute hepatitis B patients. (2) BCP mutation may impair HBeAg expression. (3) PCR microplate sandwich hybridization ELISA is a sensitive and efficient method for detecting gene mutations.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2002年第5期287-289,共3页
Chinese Journal of Infectious Diseases
关键词
微板核酸杂交
乙型肝炎病毒
C基因启动子突变
Microplate sandwich hybridization
Hepatitis B virus
Basic core promoter mutation
