乙型肝炎患者外周血单个核细胞HBV cccDNA含量的检测及其临床意义

Detection of HBV cccDNA of peripheral blood mononuclear cells in hepatitis B patients and its clinical significance

[目的]采用实时荧光定量PCR的方法检测乙型肝炎患者外周血单个核细胞中HBV cccDNA的含量,并探讨其临床意义。[方法]应用实时荧光PCR(RT-PCR)方法对随机选取的乙型肝炎患者100例和1例追踪治疗的乙肝患者的血清HBV DNA含量和外周血单个核细胞中HBV cccDNA的含量进行检测;对已确诊的慢性乙肝患者40例,肝硬化患者30例外周血单个核细胞中HBV cccDNA的含量进行检测。[结果]100例乙型肝炎患者中,HBVDNA含量<5×102copies/mL有57例,HBV cccDNA的含量均<5×102copies/mL;HBV DNA含量>5×102copies/mL有43例,HBV cccDNA的含量>5×102copies/mL有22例;HBV DNA含量>1×103copies/mL的患者有17例,在这些患者中HBV cccDNA的含量>5×102copies/mL者为13例,两者之间比例经χ2检验,差异无显著性意义,P>0.05;肝硬化患者HBV cccDNA阳性率(>5×102copies/mL)为67.7%,远远高于慢性乙肝患者的42.5%和随机乙肝人群的22.0%。各组间患者比例经χ2检验,差异有显著性意义,P<0.05。[结论]乙型肝炎患者进行外周血单个核细胞中HBV cccDNA的检测,对于抗病毒治疗效果的监测和预后判断都重要的临床意义。

[Objective] To detect HBV cccDNA levels of peripheral blood mononuclear cells in hepatitis B patients with RT-PCR,and discuss its clinical significance.[Methods] Serum and peripheral blood mononuclear cell samples were collected from 100 random patients with HBV infection,40 patients with clinical diagnosis of chronic hepatitis B,30 liver cirrhosis patients with HbsAg（＋）,and 1 acute hepatitis B patient during therapy.HBV cccDNA and HBV DNA levels were tested by RT-PCR.[Results] In the 100 patients with HBV infection,HBV DNA levels 5×102 copies/mL were 57,in which,HBV cccDNA levels were all 5×102 copies/mL;HBV DNA levels5×102 copies/mL were 43（43%）,HBV DNA levels 5×102 copies/mL were 22（22%）.In 43 HBV DNA positive（ 5×102 copies/mL） samples,HBV DNA 1×105 copies /mL（strong positive） were 17,HBV cccDNA 5 × 102 copies/mL were 13.There was not significant difference between these groups（P0.05）.On the other hand,positive rate of HBV cccDNA in cirrhosis patients＇ peripheral blood mononuclear cell samples（ 5×102 copies/mL） was 67.7%,which was much higher than that of chronic hepatitis B patients（42.5%） and hepatitis B random groups（22.0%）.There was significant difference between them（P0.05）.[Conclusion] RT-PCR detecting HBV cccDNA of peripheral blood mononuclear cells in hepatitis B patients has important clinical significance for monitoring the curative effect and determining their prognosis.