慢性乙型肝炎血清学标志、HBV DNA定量分析及HBV前C/C变异的临床研究

目的 :探讨慢性乙型肝炎患者血清学标志、HBVDNA载量及HBV前C/C变异与临床病变的关系。方法 :随机选取慢性乙型肝炎患者 2 4 6例 ,其中轻度 10 2例、中度 88例、重度 5 6例 ,分别应用ELISA、实时PCR法检测其血清中的HBV免疫学标志及HBVDNA含量。同时对HBeAg阴性而HBVDNA含量高拷贝的 3例轻度、6例中度及 8例重度患者血清 ,应用巢式PCR方法分离前C/C区全长基因 ,纯化后克隆入T载体 ,鉴定后进行序列测定。结果 :10 2例轻度、88例中度及 5 6例重度患者中血清学标志HBsAg ,HBeAg,抗 -HBc阳性模式分别为 5 3例 (5 1.9% )、4 6例(5 2 .3% )、33例 (5 8.9% ) ;HBsAg ,抗 -HBe,抗 -HBc阳性模式分别为 37例 (36 .3% )、32例 (37.5 % )、2 1例 (37.5 % ) ;单独HBsAg阳性为 8例 (7.8% )、5例 (5 .6 % )、1例 (1.7% ) ;其他模式分别为 4例、5例、1例。HBVDNA含量平均值分别为 10E 5 .5 3拷贝 /ml、10E 6 .0 3拷贝 /ml、10E 6 .5 8拷贝 /ml。HBeAg阴性而HBVDNA含量 >10E 6拷贝 /ml的病例数分别为 3例、6例、8例。统计分析表明 ,轻、中、重慢性乙肝患者间血清学标志及HBVDNA含量差异均不具统计学意义 ,而HBeAg阴性HBVDNA含量高拷贝的患者出现频率差异具有统计学意义。序列测定结果显示 17例患者血清的HBV分离株的前C/C

Clinical analysis of quantity of HBV DNA, serological indexes, and pre-C/C mutation in chronic hepatitis B patients

Objective: To explore the relationship between clinical profile and serological indexes, quantity of HBV DNA, and pre-C/C mutation of HBV in chronic hepatitis B patients. Methods: 246 HBV positive sera were randomly selected. ELISA and real-time-PCR were used respectively to examine the serological indexes and quantity of HBV DNA. Nest-PCR cloning was used to isolate the full-length gene of pre-C/C. Then the gene fragment was cloned into the plasmid of T-vector and sequenced. Results: In the 102 mild, 88 moderate and 56 severe hepatitis patients, the number of cases with positive HBsAg, HBeAg and anti-HBc was 53 (51.9%), 46(52.3%) and 33( 58.9%)respectively; the number of cases with positive HBsAg, anti-HBe and anti-HBc was 37 (36.3%), 32(37.5%) and 21(37.5%)respectively; the number of cases with positive HBsAg only was 8 ( 7.8%), 5(5.6%) and 1(1.7%)respectively; the mean content of HBV DNA was 10 E 5.53, 10 E 6.03 and 10 E 6.58 copy/ml respectively; the number of cases with negative HBeAg and HBV DNA>10 E 6 copy/ml was 3, 6 and 8 respectively. The differences in the serological items and HBV DNA content among mild, moderate and serious hepatitis patients were not significant, but the significant difference was found in the high HBV DNA content of HBeAg negative patients. DNA sequencing results showed that there was sequence mutation in the pre C/C region of 17 patients. The rate of mutation of A83(1 896 nt)was highest(11 patients), followed by T1862(7 patients), and A1899 and T1858 respectively. One severe patient was found having 2 083-2 203 nt absent mutation. Conclusions: The differences in serological items and HBV DNA content among the mild, moderate and severe hepatitis patients are not significant. Test for serological items can not replace the test for HBV DNA quantity, especially when HBeAg turns negative. There are relations between viral mutation and the course progression of HBV infection.