套式聚合酶链反应法检测外周血单个核细胞中HBcVccDNA

Establishment of a nested polymerase chain reaction for detection of HBV cccDNA in peripheral blood mononuclear cells

目的:建立套式聚合酶链反应法(nPCR)检测慢性乙型肝炎患者外周血单个核细胞(PBMC)中乙型肝炎病毒共价闭合环状DNA(HBV cccDNA)。方法:根据HBV基因组DNA正负链上下游缺口序列,设计两套相对保守的引物,建立HBV cccDNA套式PCR,并用该法检测200例慢性乙型肝炎患者PBMC中HBV cccDNA。结果:该法检测200例慢性乙型肝炎患者,HBV cccDNA阳性率为60%。结论:套式PCR法简便、灵敏,可用于测定PBMC中HBV cccDNA。

Objective：To establish a nested polymerase chain reaction for detection of HBV cccDNA in peripheral blood mononuclear cells. Methods：A nested polymerase chain reaction （nPCR） for detection of HBV cccDNA in peripheral blood mononuclear cells was established using two pairs of relatively conservative primers designed according to the sequences between the gap of upstream and downstream of plus and minus strand of HBV viral genomic DNA. Results： Of 200 patients with chronic hepatitis B, 120（60% ） were HBV cccDNA positive by the nPCR. Conclusion： The rPCR is simple and relatively sensitive and can be used for detection of HBV cceDNA in PBMC.