TT病毒感染者外周血单个核细胞中TT病毒的研究

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

目的 了解TTV阳性肝炎患者外周血单个核细胞 (PBMC)TTVDNA存在及复制情况。方法 采用巢式PCR(nPCR)技术和绿豆芽核酸酶消化单链DNA方法研究在PBMC中的TTVDNA存在及复制情况 ,并克隆测序分析证实。 16例血标本 (血清和PBMC)和粪便标本来自北京地坛医院1997年 10月～ 1998年 10月住院患者。结果 16例TTV阳性肝炎患者PBMC中有 14例TTVDNA阳性 ,粪便中有 5例TTVDNA阳性 ,并用绿豆芽核酸酶消化提取的 14例TTVDNA阳性患者PBMCDNA ,有 2例为TTVDNA阳性 ,提示TTVDNA能在PBMC中存在和感染。 5例非甲 庚型肝炎患者 (BDH1 5 )的血清、PBMC和粪便中TTVDNA片段部分序列经克隆测序 ,同一患者不同标本 (血清、PBMC和粪便 )序列未见差异 ,与日本株和中国株相应位置核苷酸序列同源性大于 95 % ,5例患者序列之间核苷酸同源性在 95 %～ 99%。粪便nPCR阳性患者血清病毒量明显大于粪便nPCR阴性患者。结论 TTVDNA可在TTV阳性肝炎患者PBMC中存在 。

Objective TostudythepresenceandreplicationofTTVDNAintheperipheralblood mononuclearcells (PBMC)fromhepatitispatientswithTTVinfection Methods Nested PCR (nPCR)was performedfordetectingtheTTVDNAfromthesera ,PBMCsandfecesof16patientswithnon Atonon Ghepatitis inBeijingDitanHospitalfromOct.1997toOct.1998.PBMCDNAswereaslodigestedbymungbeannuclease andamplifiedbynPCRtoclasifyifTTVDNAwasstillpositive nPCRproductsofthesera ,PBMCsandfecesof5 hepatitispatientswithnon Atonon Ghepatitiswereclonedandsequenced Results 14ofthe16samplesof PBMCand 5ofthe 16samplesofthefeceswereTTVDNApositive Twoofthe 14samplesofthePBMCwerestill positiveafterthedigestionandamplificationofthesingle standDNA Sequenceanalysisshowedthattherewas morethan 95 %nucleotidehomologyamongthesequenced 5clones (fromthesera ,PBMCandfecesofthe 5 patients)andreportedAB0 0 83 94 (fromJapan)andAF0 79173 (fromChina) Virusloadingofpatientswhose feceswerepositivebynPCRwasobviouslymorethanthepatientswithnegativeTTVDNA Conclusion TTV DNAcanexistandreplicateinthePBMC ;thehepatocytesofpatientswithTTVinfectionarenottheonlyplaceof TTVexistenceandreplication