[TT病毒感染的肝炎患者外周血单个核细胞中TT病毒DNA的存在与复制]

[Presence and replication of TTV DNA in peripheral blood mononuclear cells from hepatitis patients with TTV infection].

作者信息

He Z, Feng H, Zhou Y

机构信息

Beijing Ditan Hospital, Beijing 100011, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2000 Jan;80(1):16-9.

PMID:11798730

Abstract

OBJECTIVE

To study the presence and replication of TTV DNA in the peripheral blood mononuclear cells (PBMC) from hepatitis patients with TTV infection.

METHODS

Nested-PCR (nPCR) was performed for detecting the TTV DNA from the sera, PBMCs and feces of 16 patients with non-A to non-G hepatitis in Beijing Ditan Hospital from Oct. 1997 to Oct. 1998. PBMC DNAs were aslo digested by mung bean nuclease and amplified by nPCR to clasify if TTV DNA was still positive. nPCR products of the sera, PBMCs and feces of 5 hepatitis patients with non-A to non-G hepatitis were cloned and sequenced.

RESULTS

14 of the 16 samples of PBMC and 5 of the 16 samples of the feces were TTV DNA positive. Two of the 14 samples of the PBMC were still positive after the digestion and amplification of the single-stand DNA. Sequence analysis showed that there was more than 95% nucleotide homology among the sequenced 5 clones (from the sera, PBMC and feces of the 5 patients) and reported AB008394 (from Japan) and AF079173 (from China). Virus loading of patients whose feces were positive by nPCR was obviously more than the patients with negative TTV DNA.

CONCLUSION

TTV DNA can exist and replicate in the PBMC; the hepatocytes of patients with TTV infection are not the only place of TTV existence and replication.

摘要

目的

研究TTV感染的肝炎患者外周血单个核细胞（PBMC）中TTV DNA的存在及复制情况。

方法

采用巢式聚合酶链反应（nPCR）检测1997年10月至1998年10月在北京地坛医院就诊的16例非甲至非庚型肝炎患者血清、PBMC及粪便中的TTV DNA。用绿豆核酸酶消化PBMC DNA后再经nPCR扩增，以判断TTV DNA是否仍为阳性。对5例非甲至非庚型肝炎患者的血清、PBMC及粪便的nPCR产物进行克隆测序。

结果

16例PBMC样本中有14例、16例粪便样本中有5例TTV DNA阳性。14例PBMC样本中的2例单链DNA经消化扩增后仍为阳性。序列分析显示，测序的5个克隆（来自5例患者的血清、PBMC及粪便）与已报道的AB008394（来自日本）和AF079173（来自中国）的核苷酸同源性超过95%。nPCR检测粪便阳性患者的病毒载量明显高于TTV DNA阴性患者。