斑点杂交法检测肝病患者TTV感染状况初探
作者：张俊 赵西平 王宝菊 杨燕 李方和 杨东亮
单位：430030 武汉同济医科大学同济医院临床免疫研究室
关键词：TTV病毒；聚合酶链反应；斑点杂交
肝脏000106摘 要：目的探讨斑点杂交法在检测肝病患者TT病毒(TTV)感染状况中的应用价值。方法提抽患者血清中DNA，分别采用聚合酶链反应(PCR)及斑点杂交技术检测其中的TTV病毒DNA。结果在30份非甲～非戊型肝炎患者血清中，PCR方法检出11份为TTV阳性，检出率为36.7%。采用斑点杂交法检出3份阳性血清，阳性率为10%。制备的探针具有TTV特异性，可检出相当于106拷贝的病毒DNA。结论以地高辛标记的TTV探针用于斑点杂交可以检出血清病毒滴度较高的TTV，其敏感度低于PCR方法。 Detection of TTV DNA in sera of patients with non A～E hepatitis by dot blot hybridization
ZHANG Jun,ZHAO Xi-ping,WANG Baoju,et al.
(Research Unit of Clinical Immunology,Tongji Hospital,Tongji Medical University,Wuhan 430030)
Abstract：Objective To explore the possibility of using dot-blot hybridization in the detection of TTV virus infection.Methods Nuclear acid was extracted from serum specimens of patients with non A～E hepatitis,PCR and dot-blot hybridiza-tion were carried out respectively to detect TTV DNA.Results Of 30 sera specimens,11 were found positive by PCR,3 were found positive by dot-blot hybridization.The probe prepared was demonstrated to be TTV-specific,and the sensitivity was de-termined to be capable of detecting 106 copies of viral DNA.Conclusion Dot-blot hybridization with DIG-labelled TTV DNA probe can be employed to detect TTV DNA in sera samples containing relatively high titer virus,with a sensitivity lower than that of PCR.
Keywords：TTV;Dot-blot hybridization;PCR
参考文献：
[1]Nishizawa T,Okamoto H,Konishi K,et al.A novel DNA virus(TTV)assosicated with elevated transaminase level in posttransfusion hepatitis of unknown etiology.Biochemi Biophys Res Commun,1997,241:92-97.
[2]Pessoa MG,Terrault NT,Detmer J,et al.Quantitation of hepatitis G and C virus in 1iver:evidence that hepatitis G vires is not hepatotropic.Hepatology,1997,27:505-508.
[3]陈红松,丛旭,王宇.经输血传播病毒(TTV)在血液透析患者中感染状况及基因分析.中华肝脏病杂志,1998,6:164-166.
[4]马为民,周北平,徐六妹,等.种新的可经输血传播病毒(TTV)的分子流行病学.中华肝脏病杂志,1998,6:167-169.
[5]骆抗先,章廉,王珊珊,等.一种新型肠传性肝炎的临床流行病学的初步研究.中华肝脏病学杂志,1998,6:161-163.
收稿日期：1999-08-12, 百拇医药
单位：430030 武汉同济医科大学同济医院临床免疫研究室
关键词：TTV病毒；聚合酶链反应；斑点杂交
肝脏000106摘 要：目的探讨斑点杂交法在检测肝病患者TT病毒(TTV)感染状况中的应用价值。方法提抽患者血清中DNA，分别采用聚合酶链反应(PCR)及斑点杂交技术检测其中的TTV病毒DNA。结果在30份非甲～非戊型肝炎患者血清中，PCR方法检出11份为TTV阳性，检出率为36.7%。采用斑点杂交法检出3份阳性血清，阳性率为10%。制备的探针具有TTV特异性，可检出相当于106拷贝的病毒DNA。结论以地高辛标记的TTV探针用于斑点杂交可以检出血清病毒滴度较高的TTV，其敏感度低于PCR方法。 Detection of TTV DNA in sera of patients with non A～E hepatitis by dot blot hybridization
ZHANG Jun,ZHAO Xi-ping,WANG Baoju,et al.
(Research Unit of Clinical Immunology,Tongji Hospital,Tongji Medical University,Wuhan 430030)
Abstract：Objective To explore the possibility of using dot-blot hybridization in the detection of TTV virus infection.Methods Nuclear acid was extracted from serum specimens of patients with non A～E hepatitis,PCR and dot-blot hybridiza-tion were carried out respectively to detect TTV DNA.Results Of 30 sera specimens,11 were found positive by PCR,3 were found positive by dot-blot hybridization.The probe prepared was demonstrated to be TTV-specific,and the sensitivity was de-termined to be capable of detecting 106 copies of viral DNA.Conclusion Dot-blot hybridization with DIG-labelled TTV DNA probe can be employed to detect TTV DNA in sera samples containing relatively high titer virus,with a sensitivity lower than that of PCR.
Keywords：TTV;Dot-blot hybridization;PCR
参考文献：
[1]Nishizawa T,Okamoto H,Konishi K,et al.A novel DNA virus(TTV)assosicated with elevated transaminase level in posttransfusion hepatitis of unknown etiology.Biochemi Biophys Res Commun,1997,241:92-97.
[2]Pessoa MG,Terrault NT,Detmer J,et al.Quantitation of hepatitis G and C virus in 1iver:evidence that hepatitis G vires is not hepatotropic.Hepatology,1997,27:505-508.
[3]陈红松,丛旭,王宇.经输血传播病毒(TTV)在血液透析患者中感染状况及基因分析.中华肝脏病杂志,1998,6:164-166.
[4]马为民,周北平,徐六妹,等.种新的可经输血传播病毒(TTV)的分子流行病学.中华肝脏病杂志,1998,6:167-169.
[5]骆抗先,章廉,王珊珊,等.一种新型肠传性肝炎的临床流行病学的初步研究.中华肝脏病学杂志,1998,6:161-163.
收稿日期：1999-08-12, 百拇医药