上海地区血液透析患者与供血者中输血传播病毒DNA检测及部分核苷酸序列分？

目的 了解上海地区暴露于血及血制品的血透患者中输血传播病毒（ＴＴＶ）感染。方法 采用套式ＰＣＲ技术，检测了６例血透患者和４９例供血员血清中ＴＴＶ ＤＮＡ，并对其中各１例ＰＣＲ产物（２７２ｂｐ）测序。结果 血透患者ＴＴＶ ＤＮＡ的检出率为３２．８％（２０／６１），供务员的检出主继２４．５％（１２／４９）。２０例ＴＴＶＤＮＡ阳性血透患者中，６例为单纯ＴＴＶ感染，６例为ＴＴＶ、ＨＣＶ及ＨＧＶ重叠感染，４     

维持性血透患者六种肝炎病毒感染的调查研究

目的：了解维持性血液透析患者六种肝炎病毒感染情况,探讨其与输血、透析时间、肝功能的关系。方法：对１６７例维持性血液透析患者进行调查,采用第二代酶联免疫法（ＥＬＩＳＡ）检测ＨＡＶ、ＨＢＶ、ＨＥＶ、ＨＧ,用套式ＰＣＲ法检测经输血传播的肝炎病毒（ＴＴＶ）ＤＮＡ,并按血透时间、输血次数等分组进行对比分析。结果：ＨＡＶ、ＨＢＶ、ＨＣＶ、ＨＨＥＶ、ＨＧＶ、ＴＴＶ感染率分别为０％、２０．４％、５５．７％、０％、     

多重和套式聚合酶链反应检测血液,腹膜透析患者血清中HBV D…

利用免疫学和聚合反应（ＰＣＲ）方法，对３５例血液透析（血透），１４例腹膜透析（腹透）患者的８４份血清进行了检测。血透组抗－ＨＣＶ阳性率８２．９％，ＨＣＶ ＲＮＡ ＰＣＲ阳性率５１．４％；腹透组抗－ＨＣＶ阳性率仅７．１％，ＨＣＶ ＲＮＡ ＰＣＲ均阴性。ＨＢｓＡｇ和（或）ＨＢｅＡｇ在二组的阳性率分别为２５．７％和２１．４％；ＨＢＶＤＮＡＰＣＲ阳性率分别为４５．７％和６４．２％。透析患者ＨＣＶ和ＨＢＶ感     

多重和套式聚合酶链反应检测血液、腹膜透析患者血清中HBV DNA和HCV RNA

利用免疫学和聚合酶链反应（ＰＣＲ）方法，对３５例血液透析（血透）、１４例腹膜透析（腹透）患者的８４份血清进行了检测。血透组抗－ＨＣＶ阳性率８２．９％，ＨＣＶＲＮＡＰＣＲ阳性率５１．４％；腹透组抗－ＨＣＶ阳性率仅７．１％，ＨＣＶＲＮＡＰＣＲ均阴性。ＨＢｓＡｇ和（或）ＨＢｅＡｇ在二组的阳性率分别为２５．７％和２１．４％；ＨＢＶＤＮＡＰＣＲ阳性率分别为４５．７％和６４．２％。透析患者ＨＣＶ和ＨＢＶ感染率显著高于一般人群。血、腹透两组ＨＣＶ感染率相差非常显著，血透患者抗－ＨＣＶ和ＨＣＶＲＮＡＰＣＲ阳性高于腹透患者，表明血透过程在丙型肝炎传播方面起重要作用。作者还采用多重和套式ＰＣＲ方法，将ＨＢＶＤＮＡ和ＨＣＶＲＮＡ在合并的逆转录和ＰＣＲ扩增系统中，连续进行逆转录和第一轮多重ＰＣＲ扩增，再进行第二轮多重ＰＣＲ，具有特异、敏感、简便、快速和经济等特点，适于临床应用。     

一种新型肝炎相关性病毒的分子生物学研究

目的 了解南京地区ＴＴＶ病毒感染情况，重叠ＴＴＶ感染对慢性乙型肝炎病变和蔼和ＨＢＶ复制的影响。方法 采用巢式ＰＣＲ方法检测血清标本中ＴＴＶ－ＤＮＡ。结果 ４６９ 血清标本中，ＴＴＶＤＮＡ总检出率为２１．１％。其中健康人群、献血员、血透患者、甲型肝炎、乙型肝炎、丙型肝炎、戊型肝炎、庚型肝炎、非甲－非庚型肝炎检出率分别是９．４％、３３．３％、３０．８％、１１．６％、１９．９％、１５．４％、８．７％、９     

TTV感染血清学,原位杂交及干扰素治疗研究

目的了解ＴＴＶ感染状况,探讨ＴＴＶ致病性及对干扰素治疗的应答。方法采用ＰＣＲ和原位杂交方法检测血清和肝组织中ＴＴＶ ＤＮＡ,回顾分析８例ＴＴＶ／ＨＣＶ感染者干扰素治疗后病毒及ＡＬＴ应答状况。结果各型肝病患者（３６．７％～４２．８％）和职业供血员（４３．２％）ＴＴＶ感染率均显著高于义务供血员（健康对照人群,５．６％）。部分感染者采用斑点杂交方法可自血清中检出ＴＴＶ ＤＮＡ（３／３０,１０．０％）,提示存在较高的血清病毒负荷, １６例慢性非甲～戊型肝炎患者及１６例肝炎后肝硬化患者肝组织以原位杂交检测,各６例ＴＴＶ ＤＮＡ阳性（３７．５％）。 ８例ＴＴＶ／ＨＣＶ混合感染者经２４周疗程干扰素治疗后,６例维持持久ＡＬＴ应答（停药后２４周ＡＬＴ正常）,治疗１２周时, ８例血清ＴＴＶ ＤＮＡ均阴转,但 ２例停药后复阳, ＡＬＴ无应答的２例患者, ＨＣＶ ＲＮＡ呈阳性但ＴＴＶ ＤＮＡ阴怀;而ＴＴＶ ＤＮＡ阳转的２例患者ＡＬＴ均维持应答。结论 ＴＴＶ感染在肝病患者和职业供血员中较普遍, ＴＴＶ可在肝组织中活跃复制,其对干扰素治疗反应较敏感,ＴＴＶ／ＨＣＶ重叠感染时ＴＴＶ无明显致肝损伤作用。     

TTV在非甲-庚型肝炎患者中的检测、克隆及测序

目的分析ＴＴＶ（Ｔｒａｎｓｆｕｓｉｏｎｔｒａｎｓｍｉｔｅｄｖｉｒｕｓ）深圳分离株ＯＲＦ１部分基因序列。方法在ＴＴＶＯＲＦ１设计引物，建立巢式聚合酶链反应（Ｎｅｓｔｅｄ－ＰＣＲ），检测４０例广东地区非甲－庚型肝炎患者血清中ＴＴＶＤＮＡ。对ＰＣＲ产物进行分子克隆，以荧光法（ＡｐｐｌｉｅｄＢｉｏｓｙｓｔｅｍｓ，３７３Ａ）测序。结果４０例非甲－庚型肝炎中２１例ＴＴＶＤＮＡ阳性（５２．５％）。对其中一株ＴＴＶ（ＳＺ１）ＯＲＦ１部分基因克隆、测序，并与Ｏｋａｍｏｔｏ等报道的２株（Ｎ２２、Ｇ１ａ）相比较，其核苷酸序列的同源性分别为９６．７％与９７．４％。结论本研究证实我国华南地区存在ＴＴＶ感染。ＴＴＶ的分子克隆与测序及ＰＣＲ诊断技术的建立对国内进一步开展ＴＴＶ感染的诊断与流行病学调查均具有重要意义     

健康人群和肝病患者中检测TTV的意义

目的了解新型肝炎病毒－ＴＴＶ的致病性和在健康人群和肝病患者中的流行情况．方法收集１８０份健康体检患者血清和１５６份不同类型肝病患者血清,采用ＰＣＲ方法检测ＴＴＶ的ＤＮＡ．同时检测ＨＡＶ,ＨＢＶ,ＨＣＶ,ＨＥＶ和ＨＧＶ感染标志,比较分析ＴＴＶ在健康人群和不同类型肝病患者中流行情况及其致病性．结果健康体检人群和肝病患者中,ＴＴＶＤＮＡ检出率分别为２２％和４５％,两组间无显著性差异（Ｐ＞００５）．体检人群中,ＡＬＴ正常和升高者的检出率分别为１７％和１４３％．急性肝炎,慢性肝炎和肝硬变者的检出率分别为４８％,４３％和４７％．１１例阳性患者中,３例ＡＬＴ正常,８例ＡＬＴ异常．在８例ＡＬＴ异常患者中,６例为ＨＢＶ现行感染,１例为ＨＣＶ现行感染,仅１例为ＮＡ－Ｇ肝炎患者．结论在中国健康体检人群和肝病患者中能检出低水平的ＴＴＶ现行感染．但似乎仅引起个别患者的转氨酶轻度升高．ＴＴＶ的致病性可能较弱或需要其他因素协同致病．     

TTV阳性肝病患者的临床与病理学特征初步观察

目的 观察ＴＴＶ阳性肝病患者的临床和病理特征。方法 用巢式ＰＣＲ法检测ＴＴＶ－ＤＮＡ,同时观察血清生化指标,肝组织活检观察其病理变化。结果 ＴＴＶ感染者的血清学模式以重叠ＨＢＶ、ＨＣＶ、ＨＡＶ、ＨＥＶ、ＨＧＶ二重感染为主占５７．９％（２２／３８）,以乏力、纳差、腹胀为临床特征,ＴＢｉＬ、ＡＬＴ变化不明显,ＴＴＶ单独感染者为４２．１％（１６／３８）,急性发病者临床特征多与“急性黄疸性肝炎”相似,病理改变与各型病毒性肝炎的病理变化基本相似,但主要以肝门脉区炎症,小叶间胆管损伤,灶状坏死为多见。结论 ＴＴＶ可与各型已知肝炎病毒重叠感染也可单独感染。     

上海地区Ⅱ、Ⅲ型丙型肝炎病毒包膜区cDNA序列分析

目的 分析上海地区丙型肝炎病毒（ｈｅｐａｔｉｔｉｓ Ｃ ｖｉｒｕｗ,ＨＣＶ）Ⅱ、Ⅲ型包膜区ｃＤＮＡ序列变异。方法 采用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）扩增ＨＣＶ包膜区Ｅ１、Ｅ２／ＮＳ１片断（１ ００５ｂｐ）,在３７３ＤＮＡ全自动测序仪上进行了序列分析。结果 ＨＣＶ－Ｅ１、Ｅ２／ＮＳ１区核苷酸和氨基酸同源性比较显示：上海株Ⅱ型、Ⅲ型间同源性分别为６２．４８％和５９．１０％;上海株与加内外同型株间     

武汉地区职业献血员血清TTV等病毒核酸的检测

目的 调查武汉地区既往职业献血员血清TTVV DNA及其他几种肝炎病毒基因的状况与特点。方法采用PCR或巢式PCR对74例既往乡村献血员进行血清TTV DNA及HCV RNA、HGV RNA的检测。结果 TTVDNA、HCV RNA及HGV RNA阳性检出率分别为43.2％、23.0％与2.7％,TTV DNA检出显著高于其他肝炎相关病毒,HCV RNA阳性率显著高于HGV RNA阳性率。结论 既往长期的献血已造成地区乡村职业献血员TTV与HCV感染率的显著升高。     

High frequencies of HGV and TTV infections in blood donors in Hangzhou

AIM: To determine the frequencies of HGV and TTV infections in blood donors in Hangzhou. METHODS: RT-nested PCR for HGV RNA detection and semi-nested PCR for TTV DNA detection in the sera from 203 blood donors, and nucleotide sequence analysis were performed. RESULTS: Thirty-two (15.8%) and 30 (14.8%) of the 203 serum samples were positive for HGV RNA and TTV DNA, respectively. And 5 (2.5%) of the 203 serum samples were detectable for both HGV RNA and TTV DNA. Homology of the nucleotide sequences of HGV RT-nested PCR products and TTV semi-nested PCR products from 3 serum samples compared with the reported HGV and TTV sequences was 89.36%, 87.94%, 88.65% and 63.51%, 65.77% and 67.12%, respectively. CONCLUSION: The infection rates of HGV and/or TTV in blood donors are relatively high, and to establish HGV and TTV examinations to screen blood donors is needed for transfusion security. The genomic heterogeneity of TTV or HGV is present in the isolates from different areas.     

Transmission of and liver injury by TT virus in patients on maintenance hemodialysis

To study the prevalence and clinical significance of TT virus (TTV) infection in hemodialysis patients, we tested for TTV DNA in serum, using the nested polymerase chain reaction. The prevalence of TTV DNA in 352 hemodialysis patients was 32%, significantly higher than that in 50 healthy blood donors (12%). The prevalence increased with age (P = 0.0098); it was 20% (22/110) in patients aged less than 49 years, 37% (69/188) in those aged 50–69 years, and 41% (22/54) in those aged over 70 years. Other clinical features and the prevalence of other hepatitis viral markers tested did not differ between patients with TTV DNA and those without it. The detection rate of hepatitis C virus (HCV) and hepatitis G virus (HGV) viremias increased with duration of hemodialysis and with the number of blood transfusion units, but the prevalence of TTV viremia did not. Twenty-nine of 91 patients followed for 5 years were initially positive for TTV DNA. Of these 29 patients, 17 (59%) carried this viremia for at least 5 years. Fourteen of the 62 patients (23%) who were initially negative for TTV DNA acquired TTV viremia. Serum alanine aminotransferase (ALT) levels were elevated in patients with HCV viremia but not in patients with HGV or TTV viremia. However, the mean ALT level in patients with all three viremias (HCV, HGV, and TTV) was significantly higher than that in patients with one or two of the viremias. More than 30% of the hemodialysis patients had TTV viremia and the carrier state was maintained for years. The hemodialysis procedures, including blood transfusion, did not seem to be crucial for the transmission of TTV. The pathogenic effects of TTV on hepatitis appear to be limited. (Received July 21, 1998; accepted Sept. 25, 1998)     

High rate of TTV infection in multitransfused patients with pediatric malignancy and hematological disorders

The prevalence of transfusion-transmitted virus (TTV) infection has not been known in patients suffering from pediatric malignancies and hematological disorders who receive blood transfusion and/or blood products during treatment. Blood samples were taken from 75 patients. TTV infection was identified when TTV DNA was detected in serum by a polymerase chain reaction (PCR) assay. Hepatitis C virus (HCV) and hepatitis G virus (HGV) RNA were also assayed by PCR. TTV DNA was detected in 38 of 75 patients (51%). In 4 of 38 patients, the amount of blood transfused was less than 3 units. By time since last transfusion, TTV DNA was detected in 12 of 35 patients after more than 4 years, 12 of 21 between 1 and 4 years, and 14 of 19 within 1 year. Six patients had mixed infection of TTV and HCV, and 12 patients had mixed infection of TTV and HGV. Three different kinds of virus were found simultaneously in serum from 3 patients. Eight out of 75 patients showed abnormal levels of alanine aminotransferase (ALT) (>40 IU/liter), and 3 of them had TTV DNA. All patients who had TTV DNA and elevated ALT levels also were positive for HCV RNA and HGV RNA. The prevalence of TTV infection is high in patients with pediatric malignancies and hematological disorders after episodes of blood transfusion. Transfusion is one of the most important risk factors for TTV infection regardless of the amount of blood transfused.     

Investigation of HGVand TTVinfection in sera and saliva from non—hepatitis patients with oral diseases

AIM：To determine the frequencies of HGVand TTVinfections in serum and saliva samples of non-hepatitis patients with oral diseases in Hangzhou area,and to understand the correlation between detected results of HGVRNAand/orTTVDNAin sera and in saliva from the same patients.METHODS:RT-nested PCRfor HGVRNAdetection and semi-nestedPCRfor TTVDNAdetection were performed in the serum and saliva samples from226non-hepatits patients with oral diseases,and nucleotide sequence analysis.RESULTS:Twenty-seven(11.9%)and21(9.3%)of the 226serum samples were only positive for HGVRNAand TTVDNA,respectively,10(4.4%)and9(3.9%)of the 226saliva samples were only positive for HGVNA and TTVDNA,respectively.And7(3.1%)ofthe serum samples and2(0.9%)of the saliva samples showed the positive amplification results for bothHGVRNAand TTVDNA.12saliva samples from the 34patients(35.3%)withHGVor HGV/TTVviremia and 11saliva samples from the 28patients(39.3%)with TTV or HGV/TTVviremia were HGVRNAdetectable,respctively,including two patients positive for bothHGVRNAand TTVDNAin serum and saliva samples.No saliva samples from the 226patients were found to be HGVRNAor TTVDNAdetectable while their serum samples were negative for HGVorTTV.Homologies of the nucleotide sequences of HGVand TTVamplification products from the serum and saliva samples of the two patients compared with the reported sequences were 88.65±91.49%and65.32-66.67%,respectively,In comparison with the nucleotide sequences of amplification products between serun and fromsaliva sample from any one of the two patients,the homlogies were98.58%and 99.29%for HGV,and were98.65%and98.20%forTTV,respectively.CONCLUSION:Relatively high carrying rates of HGVand/orTTVin the sera of non-hepatitis patients with oral diseases in Hangzhou area are demonstrated.Parts of the carriers are HGVand/orTTVpositive in their saliva.The results of this study indicate that dentists may be one of the populations with high risk for HGVand/orTTVinfection.and by way of saliva HGV and TTVmay be transmitted among individuals.     

G Virus in the General Population and in Patients on Hemodialysis

To determine the routes of transmission ofhepatitis G virus (HGV) and the relationship between HGVand hepatitis C virus (HCV) infections, we tested forHGV RNA by polymerase chain reaction and antibody to HCV (anti-HCV) in 494 hemodialysis patients,638 inhabitants of two HCV endemic areas, and in 400blood donors in Japan. HGV RNA was detected in 6.9% ofhemodialysis patients, in 1.4% of inhabitants, and in 0.8% of donors, and anti-HCV wasdetected in 39.3%, 12.4%, and 1.8%, respectively. Of HGVRNA-positive hemodialysis patients, and HGV RNA-positiveinhabitants, 64.7% and 11.1%, respectively, had been given blood transfusions. Theprevalences of HGV RNA and anti-HCV significantlyincreased with the duration of hemodialysis. Of all HGVRNA positives, 74.4% were coinfected with HCV andsubjects with HGV RNA alone had normal liver function.In conclusion, HGV is transmitted by blood transfusionand within the hemodialysis unit itself. HGV does notseem to injure hepatocytes.     

Infection with TT virus, a novel transfusion-transmissible DNA virus, in haemophiliacs and in blood products

We evaluated the characteristics and rate of infection with TT virus (TTV), a novel DNA virus, in Japanese haemophiliacs. TTV DNA was measured in 60 haemophiliacs by semi-nested polymerase chain reaction. Co-infection with hepatitis C virus (HCV), hepatitis G virus (HGV) and human immunodeficiency virus (HIV) was also evaluated. In addition, the rate of detection of TTV DNA in blood products was evaluated. TTV DNA was detected in 35/60 haemophiliacs (58.3%). There were no differences in the backgrounds or characteristics between haemophiliacs with and without TTV infection, except for higher levels of IgG and IgM in patients with TTV infection. In patients infected with TTV of types other than type 1, which are rarely detected in Japan, the rate of co-infection with HCV of imported types was high; TTV of types other than type 1 in Japanese haemophiliacs were probably transmitted by imported blood products. TTV DNA was detected in over half of the blood products tested, but TTV DNA concentrations in these products were lower than in the serum of haemophiliacs.     

TT virus and hepatitis G virus infections in Korean blood donors and patients with chronic liver disease

AIM: To determine the prevalences of TTV and HGV infections among blood donors and patients with chronic liver disease in Korea, to investigate the association of TTV and HGV infections with blood transfusion, and to assess the correlation between TTV and HGV viremia and hepatic damage. METHODS: A total of 391 serum samples were examined in this study. Samples were obtained from healthy blood donors (n=110), hepatitis B surface antigen (HBsAg)-positive donors (n=112), anti-hepatitis C virus (anti-HCV)-positive donors (n=69), patients with type B chronic liver disease (n=81), and patients with type C chronic liver disease (n=19). TTV DNA was detected using the hemi-nested PCR. HGV RNA was tested using RT-PCR. A history of blood transfusion and serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also determined. RESULTS: TTV DNA was detected in 8.2 % of healthy blood donors, 16.1 % of HBsAg-positive donors, 20.3 % of anti-HCV-positive donors, 21.0 % of patients with type B chronic liver disease, and 21.1 % of patients with type C chronic liver disease. HGV RNA was detected in 1.8 % of healthy blood donors, 1.8 % of HBsAg-positive donors, 17.4 % of anti-HCV-positive donors, 13.6 % of patients with type B chronic liver disease, and 10.5 % of patients with type C chronic liver disease. The prevalence of TTV and HGV infections in HBV- or HCV-positive donors and patients was significantly higher than in healthy blood donors (P<0.05), except for the detection rate of HGV in HBsAg-positive donors which was the same as for healthy donors. There was a history of transfusion in 66.7 % of TTV DNA-positive patients and 76.9 % of HGV RNA-positive patients (P<0.05). No significant increase in serum ALT and AST was detected in the TTV- or HGV-positive donors and patients. CONCLUSION: TTV and HGV infections are more frequently found in donors and patients infected with HBV or HCV than in healthy blood donors. However, there is no significant association between TTV or HGV infections and liver injury.     

重庆地区HGV感染的分子流行病学

目的 探讨重庆地区HGV感染和基因型特征，了解致病性和传播途径。方法 用RT-PCR和ELlSA方法检测685例献血员和76例血液透析患者HGV感染状况，比较肝功能和重叠感染情况并进行部分病例随访；进行HGV 5-NCR测序。结果 本地存在HGV感染．血液透析患者HGV RNA阳性率(36％)明显高于献血员抗-HGV阳性率(3％)，约半数透析患者HGV合并HCV和HBV感染；基因分型表明属于第3组3b亚型。结论 HGV主要经血传播，感染HGV透析患者未发现有明显致病性；基因分型有助于深入探讨病毒致病性和变异。