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目的分析乙型肝炎(乙肝)病毒e抗原(HBeAg)阴性慢性乙型肝炎(chronic hepatitis B,CHB)患者血清乙肝病毒DNA(HBV DNA)、乙肝病毒表面抗原(HBsAg)和乙肝病毒cccDNA的相关性,探讨CHB患者肝细胞内外与活动性肝炎有关的病毒学转变过程。方法对南京第二医院64例未经治疗的HBeAg阴性的CHB患者均进行了肝穿刺获取肝组织样本。按照丙氨酸氨基转移酶>80U/L,HBV DNA>100 000拷贝/ml诊断为活动性肝炎,否则为非活动性肝炎的标准分为两组。化学发光法检测血清HBsAg,Taqman实时PCR法定量检测血清及肝组织中HBV DNA含量,同时检测肝组织中cccDNA。结果血清HBV DNA和血清cccDNA明显相关(r=0.483,P<0.01),和lg肝细胞内总cccDNA明显相关(r=0.723,P<0.01)。血清HBsAg和cccDNA及lg肝细胞内HBV DNA均没有明显的相关性。活动性HBeAg阴性的CHB患者的lg cccDNA及lg肝细胞内总HBV DNA,明显高于非活动性HBeAg阴性的CHB患者。活动性HBeAg阴性的CHB患者,乙肝病毒的复制效率(血清HBV DNA和cccDNA的比率)高出非活动性CHB患者20%以上。血清HBsAg水平及HBsAg与cccDNA的比率在两组患者间差异均无统计学意义。结论 HBeAg阴性的CHB患者,血清HBV DNA水平和cccDNA有明显的相关性,血清HBV DNA可以作为肝细胞内cccDNA含量的替代指标,代表着更高效的HBV复制效率。
Objective To analyze the correlation between serum HBV DNA, HBsAg and HBV cccDNA in patients with chronic hepatitis B (CHB) To investigate the virological transformation process of active hepatitis in hepatocytes of patients with CHB. Methods Sixty-six patients with untreated HBeAg-negative CHB in Nanjing Second Hospital were enrolled in this study. In accordance with the alanine aminotransferase> 80U / L, HBV DNA> 100000 copies / ml diagnosis of active hepatitis, otherwise the standard of inactive hepatitis is divided into two groups. The chemiluminescence method was used to detect the serum HBsAg, and the Taqman real-time PCR method was used to quantitatively detect the HBV DNA content in serum and liver tissue. At the same time, the cccDNA in liver tissue was detected. Results Serum HBV DNA was significantly associated with serum cccDNA (r = 0.483, P <0.01), and significantly correlated with total cccDNA in lg hepatocytes (r = 0.723, P <0.01). Serum HBsAg and cccDNA and lg hepatocyte HBV DNA were not significantly correlated. Lg cccDNA in active HBeAg-negative CHB patients and total HBV DNA in 1g hepatocytes were significantly higher than those in non-active HBeAg-negative CHB patients. In patients with active HBeAg-negative CHB, HBV replication efficiency (serum HBV DNA and cccDNA ratio) is higher than 20% of inactive CHB patients. Serum HBsAg levels and the ratio of HBsAg to cccDNA were not significantly different between the two groups. Conclusions Serum HBV DNA levels are significantly correlated with cccDNA in patients with HBeAg-negative CHB. Serum HBV DNA can be used as a surrogate marker for the content of cccDNA in hepatocytes, which represents a more efficient HBV replication efficiency.