摘要
目的通过对外周血单个核细胞（PBMCs）中乙型肝炎病毒（HBV）DNA和HBV共价闭合环状DNA（cccDNA）的检测，证实不同临床类型乙型肝炎患者PBMCs被HBV感染的事实。方法提取PGM-HBV质粒，用双蒸水连续10倍梯度稀释，建立浓度为3×10^2～3×10^5拷贝／mL PGM-HBV标准系列；常规分离全血中的PB-MCs，提取DNA，应用实时荧光定量聚合酶链反应法检测PBMCs中HBV DNA和HBVcccDNA。结果78例慢性乙型肝炎患者，PBMCs中HBVcccDNA阳性31例（39．74％）。HBVcccDNA与血清和PBMCs中HBV DNA阳性率和滴度有良好的一致性。但在不同临床类型的乙型肝炎患者之间，PBMCs中HBVcccDNA的检出率无差别（P〉0．05）。结论根据HBVcccDNA检测结果阳性，进一步证实HBV能感染PBMCs，并在其中复制；PBMCs中HBVcccDNA阳性与疾病的炎症活动和疾病的严重性无关。
Objective To identify the evidences that HBV can infect peripheral blood monouclear cells（PBMCs） of hepatitis B patients with different clinical types by detection of HBV DNA and HBV covalently closed circular DNA （HBVcccDNA） in PBMCs. Methods Plasmid PGM-HBV, as a positive HBVcccDNA templet control, was extracted and diluted 10-fold serially from 3×10^2 copies/mL to 3 ×10^5 copies/mL with double distilled water. After PBMCs were routinely isolated from whole blood, PBMCs DNA was extracted and both HBV DNA and HBVcccDNA were detected by real-time fluorescent quantitative polymerase chain reaction. Results There was 31 cases（39. 74%） with positive HBVcccDNA in PBMCs of 78 patients with chronic hepatitis B, and the titers and positive rates of HBVcccDNA in PBMCs had a good correspondence with that of serum and PBMCs HBV DNA detection. There was no ally difference in positive HBVcccDNA of PBMCs between hepatitis B patients with different clinical situations（P〉0. 05）, Coclusion Presence and replication status of HBV in PBMCs are revealed by HBVccc DNA detection. The positive of HBVcccDNA in PBMCs is not correlated with the inflammatory activity and disease progression of chronic hepatitis B.
出处
《中国感染控制杂志》
CAS
2007年第5期292-296,共5页
Chinese Journal of Infection Control
关键词
肝炎病毒
乙型
共价闭合环状DNA
外周血单个核细胞
肝炎
乙型
慢性
hepatitis B virus
HBV covalently closed circular DNA
peripheral blood mononuclear cells
chronic hepatitis B
