体内与转染细胞中乙型肝炎病毒株复制特性的相关性

目的 比较不同乙型肝炎病毒（HBV）株在体内及转染细胞中复制特性是否相符。方法 以核酸杂交定量和聚合酶链反应-酶联免疫吸附试验（PCR-ELISA）测定5例孕妇血清中HBVDNA含量，并测定乙型肝炎表面抗原（HBsAg)和乙型肝炎e抗原（HBeAg)含量，分别克隆血清中的HBV基因组转染细胞，检测培养上清液中HBsAg和HBeAg表达水平，并以Southern印迹及核酸杂交检测转染细胞内、外HBVDNA复制水平。结果 转染细胞内、外HBVDNA复制水平与相应血清HBVDNA量呈正相关趋势，转染细胞表达的病毒抗原水平与相应血清中病毒抗原含量也呈正相关趋势。结论 感染者血清中HBVDNA和病毒抗原的含量与毒株在细胞中复制和抗原表达水平有相符趋势。HBV不同毒株在转染细胞中的复制可基本反映体内毒株的复制特性。

Correlation of replication efficiency and antigen expression of HBV strains in vivo and in transfected cells

Objective To study the correlation of replication efficiency and antigen expression of hepatitis B virus (HBV) strains in patient sera and transfected cells. Methods Quantification of five maternal serum HBV DNA was carried out using dot hybridization and polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA), hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) levels in the sera were determined by ELISA. Full-length genomes of HBV strains cloned from five pregnant women were separately used to transfect HepG2 cells. The levels of HBsAg and HBeAg in the supernatant of transfected cells were determined with Abbott EIA kits, the replication efficiency of intracelluar replicative intermediates and extracellular HBV DNA of viral particles was analyzed by Southern blot and hybridization. Results Intracellular and extracellular HBV replication and antigen expression of cloned HBV strains showed positive correlation tendency with the HBV DNA and antigen levels in serum samples. Conclusions Virus replication efficiency and expression of HBsAg and HBeAg by full-length HBV clones are in relative coincidence with the biological characteristics of HBV strains in patients. Cell transfection can be used to study the biological characteristics of individual isolates.