套式PCR和原位杂交技术检测肝病患者单个核细胞内TTV DNA

Detection of TT virus DNA in peripheral blood mononuclear cells in patients with chronic hepatitis B and hepatocellular carcinoma using PCR and in situ hybridization

目的 了解慢性乙型肝炎(中度)和原发性肝癌(HBsAg阳性)患者PBMC内TTVDNA存在情况。方法 采用套式PCR以及原位杂交技术检测外周血单个核细胞(PBMC)内TTVDNA。结果 套式PCR检测26例慢性乙型肝炎(中度)患者PBMC内TTVDNA阳性7例,阳性率269%,非常显著高于健康对照(χ2=143,P<0.001);21例原发性肝癌(HBsAg阳性)患者PBMC内TTVDNA阳性4例,阳性率190%,显著高于健康对照(χ2=486,P<0.05)。7例PBMC内TTVDNA阳性的慢性乙型肝炎(中度)患者PBMC原位杂交,其中4例细胞内阳性。结论 慢性乙型肝炎(中度)和原发性肝癌(HBsAg阳性)患者PBMC细胞浆内存在TTVDNA,且阳性率相当高。

Objective To determine whether the TTV infection occurs in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis B (of medium degree) and in HBsAg positive hepatocellular carcinoma (HCC).Methods TTV DNA in PBMCs was detected by nested polymerase chain reaction(PCR) and in situ hybridization (ISH).Results 7 of 26 cases of chronic hepatitis B were positive for TTV DNA by nested PCR with a positive rate of 26.9% that was significantly higher than that of normal control(χ 2=14.3, P <0.001); 4 of 21 HBsAg positive cases were positive for TTV DNA by nested PCR with a positive rate of 19.0%, also significantly higher than that of normal control(χ 2=4.86, P <0.05).4 of 7 TTV positive patients with chronic hepatitis B tested by nested PCR were positive for TTV DNA by ISH in PBMC cytoplasm, with a positive rate of 57.1%.Conclusion (1)A higher incidence of TTV DNA in PBMCs of patients with chronic hepatitis B and in HBsAg positive HCC was identified by nested PCR and ISH; (2)TTV DNA was in the cytoplasm of PBMCs in patients with chronic hepatitis B.