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.     

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.     

A high frequency of GBV-C/HGV coinfection in hepatitis C patients in Germany

AIM:To detect infection rate of GBV-C/HGV in hepatitis C patients, to determine the methods of higher sensitivity and the primers of higher efficiency for GBV-C/HGV RNA detection and to study the dominant subtype and mutation of GBV-C/HGV.METHODS:Quantitative RT-PCR for detection pf HCV RNA concentration in serum samples, RT-nested PCR with two sets of primers for detection of GBV-C RNA, RT-PCR ELISA with two sets of primers for detection of HGV RNA, nucleotide sequence and putative amino acid sequence analysis.RESULTS:The positive rates of GBV-C RNA at the 5'-NCR and NS3 region in 211 serums amples from the patients with HCV infection were 31.8% and 22.8% respectively. The positive rates of HGV RNA at the 5'-NCR and NS5 region in the same samples were 47.9% and 31.8% respectively. The total positive rate of GBV-C/HGV RNA was as high as 55.5%. HCV copy numbers in the patients without GBV-C/HGV coinfection were statistically higher than that in the patients with GBV-C/HGV coinfection (P<0.01).Frequent mutation of nucleotide residue was present in the amplification products. Frameshift mutation was found in two samples with GBV-C NS3 region nucleotide sequences. All nucleotide sequences from amplification products showed higher homology to HGV genome than to GBV-C genome even though part of the sequences were amplified with GBV-C primers.CONCLUSION:A high frequency of GBV-C/HGV coinfection existed in the hepatitis C patients. RT-PCR ELISA was more sensitive than RT-nested PCR for detection of GBV-C/HGV RNA. The primers derived from the 5'-NCR was more efficient than those derived from the NS3 and NS5 regions. A reverse relationship was found to exist between HCV RNA concentration and GBV-C/HGV infection frequency. HGV was the dominant subtype of the virus in the local area. The major mutations of GBV-C/HGV genomes were random mutation of nucleotide residue.     

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。     

原发性肝癌患者血清和肝组织中庚型肝炎病毒基因的PCR检测

用逆转录－聚合酶链反应法（ＲＴ－ＰＣＲ）检测了原发性肝癌（ＰＬＣ）患者血清及肝癌和癌旁肝组织中的庚型肝炎病毒（ＨＧＶ）ＲＮＡ，以ＰＣＲ－双脱氧末端终止法测定了ＰＣＲ产物的核苷酸序列。结果显示，血清和肝组织中ＨＧＶＲＮＡ的检出率分别为１９．４％（１３／６７）和２５．７％（９／３５），且ＨＧＶＲＮＡ在肝癌组织呼吕旁肝组织中同时存在，血清和肝组织中ＨＧＶ ＲＮＡ检测结果的符合率为８５％；５′非编码区（５     

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。     

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)     

Transfusion-transmissible virus and hepatocellular carcinoma: a case-control study

Although transfusion-transmissible virus (TTV) is often present in the serum of patients with acute and chronic non-A–C liver diseases, its hepatotropism, pathogenicity to the liver and hepatocarcinogenicity have not been proven. We used a case-control format to compare the prevalence of TTV infection among 148 southern African Blacks with hepatocellular carcinoma and 148 matched hospital-based controls, and to test for possible interactive effects between this virus and hepatitis B virus (HBV) and hepatitis C virus (HCV) in the development of the tumour. We also determined the prevalence of TTV in 988 blood donors in Gauteng province of South Africa. The presence of TTV DNA in serum samples was detected by using the polymerase chain reaction, Southern hybridization and nucleotide sequencing. Individuals infected with TTV did not have an increased risk of developing hepatocellular carcinoma (relative risk 1.1; 95% confidence limits 0.5–2.4). Moreover, co-infection with TTV did not further increase the risk of tumour development in patients chronically infected with HBV and/or HCV. TTV was present in the serum of 2.2% of blood donors: 4.0% in Black and 1.5% in White donors. We conclude that TTV is unrelated to the development of hepatocellular carcinoma in Black Africans.     

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析

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）感染率。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］３。结果血透患者ＴＴＶ ＤＮＡ的检出率为 ３２．８％（２０／６１）,供血员的检出率为 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例为ＴＴＶ、ＨＣＶ和４例为ＴＴＶ、ＨＧＶ重叠感染。在所有ＴＴＶ感染者中,仅发现１例血清ＡＬＴ升高,该病例为ＴＴＶ、ＨＣＶ及ＨＧＶ重叠感染。对ＰＣＲ阳性扩增产物２７２ｂｐ测序结果显示,上海株（ＳＨＰ、ＳＨＤ）核苷酸的同源性为９９．６％,与深圳株、２株日本株核着酸的同源性分别为９７．４％、９８．７％和９８．７％,表明ＴＴＶ上海株与深圳株及日本株（Ｎ２２、Ｇ１ａ）属同一亚型,首次证实了上海地区的ＴＴＶ感染。结论ＴＴＶ感染可经血传播。其致病性较弱,对ＴＴＶ的研究尚属开始,ＴＴＶ的病原学、流行病学及临床意义等问题还有待进一步探索和阐明。     

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.     

Effects of HAART on hepatitis C, hepatitis G, and TT virus in multiply coinfected HIV-positive patients with haemophilia

In multiply coinfected human immunodeficiency virus (HIV)-positive patients, we investigated the effects of high-activity antiretroviral therapy (HAART) using HIV protease inhibitors on three other viruses: hepatitis C virus (HCV), hepatitis G virus (HGV), and TT virus (TTV). Viral concentrations were measured serially by polymerase chain reaction methods in five patients with quadruple infection (HIV, HCV, HGV, and TTV) and in two patients with triple infection (HIV, HCV, and HGV) before and during HAART. In addition, CD4+ cell counts and serum alanine aminotransferase (ALT) levels were measured serially. Generally we observed no difference in serum HCV RNA, HGV RNA, or TTV DNA concentrations between samples obtained before and after initiation of HAART, whereas HIV RNA concentration decreased and CD4 counts increased in most patients. However, two patients had markedly decreased concentrations of HCV RNA and HGV RNA, respectively, more than 12 months after beginning HAART. Normalization of serum ALT levels was observed in a patient with decline of HCV RNA concentrations. No interactions were observed among these four viruses. HAART had no apparent direct effects on HCV, HGV, or TTV. Further studies will be required to elucidate whether the restoration of immune status through suppression of HIV replication by HAART may affect HCV or HGV RNA concentrations.     

糖尿病患者庚型肝炎病毒感染及其临床意义

目的　了解糖尿病患者 (I型和II型 )庚型肝炎病毒 (HGV)感染情况及临床意义。方法　应用逆转录套式聚合酶链反应 (RT -NestedPCR)检测 183例糖尿病患者血清标本中的HGVRNA ,部分扩增产物进行核苷酸序列分析 ,所有患者均进行ALT检查。结果　I型和II型糖尿病患者血清标本中HGV检出阳性率分别为 4 6 7% (7/ 15 )和 2 0 2 % (34/ 16 8) ,总阳性率为 2 2 4 % (41/ 183) ;5例HGV 5’ -NCR和NS3区扩增产物与文献报道的核苷酸序列同源性分别为 87 2 %～ 89 4 %和 79 8%～ 83 5 % ;HGV感染的糖尿病患者ALT异常 (5 2～ 88U)的比例 (7 1% ,13/ 183)明显高于未感染HGV的糖尿病患者 (1 1% ,2 / 183)。结论　糖尿病患者 ,尤其是I型糖尿病患者的HGV感染率较高 ;不同地区检出的HGV基因组RNA片段的核苷酸序列存在一定的差异 ;HGV感染可导致部分患者轻微的肝脏损害。     

TT viral infection through blood transfusion: retrospective investigation on patients in a prospective study of post-transfusion hepatitis

AIM To investigate the role of blood transfusion in TT viral infection (TTV).METHODS We retrospectively studied serum samples from 192 transfusion recipients who underwent cardiovascular surgery and blood transfusion between July 1991 and June 1992. All patients had a follow-up every other week for at least 6 months after transfusion. Eighty recipients recipents blood before screening donors for hepatitis C antibody (anti-HCV), and 112 recipients reveiver screened blood.Recipients with alanine aminotransferase level ＞ 2.5 times the upper normal limit were tested for serological markers for viral hepatitis A, B,C, G, Epstein-Barr virus and cytomegalovirus.TTV infection was defined by the positivity for serum TTV DNA using the polymerase chain reaction method. RESULTS Eleven and three patients, who reveiver anti-HCV unscreened and screened blood, respectively, had serum ALT levels ＞90 IU/L. Five patients (HCV and TTV: 1; HCV,HGV, and TTV: 1; TTV: 2; and CMV and TTV: 1 )were positive for TTV DNA, and four of them had sero-conversion of TTV DNA. CONCLUSION TTV can be transmitted via blood transfusion. Two recipients infected by TTV alone may be associated with the hepatitis.However, whether TTV was the causal agent remains unsettled, and further studies are necessary to define the role of TTV infection in chronic hepatitis.     

TT virus infection in Thailand: prevalence in blood donors and patients with aplastic anemia

Aplastic anemia has been reported to occur after viral hepatitis of unknown etiology. Recently, TT virus (TTV), a novel DNA virus, was identified in a Japanese patient with posttransfusion non-A-E hepatitis. The prevalence of TTV infection was investigated among blood donors and patients with aplastic anemia in Thailand. Of 99 blood samples from blood donors, 37 tested positive for TTV DNA via semi-nested polymerase chain reaction (PCR) using TTV-specific primers. Seventeen percent of samples from blood donors younger than 20 were positive for TTV DNA, whereas 48% from donors older than 20 were positive. The high prevalence of TTV infection in Thailand is comparable to that reported in China (28%), Mongolia (43%), and Egypt (29%). Forty-two percent of newly diagnosed aplastic anemia patients tested also had TTV DNA in blood. The detection rate of TTV DNA in aplastic anemia patients does not differ significantly from rates in normal blood donors. Our present data thus argue against the role of this novel hepatitis-associated virus in the pathogenesis of aplastic anemia in Thailand. However, larger epidemiological studies may be needed to further evaluate their association.     

长春地区TT病毒的基因分型及其临床意义

为调查各种急、慢性肝炎中TT病毒的感染状况及临床意义,并检测TTV基因的分型。利用半套式PCR（semi-nested PCR）方法检测了TTV-DNA。利用邻近丁（neighbor-joining）法画出系统树。TTV-DNA的阳性率在非甲非乙非丙型急性肝炎中为42.3%,在非乙非丙型慢性肝炎中为45.5%。基因型可分为1a、1b、2a、2b等型。TTV-DNA在非甲非乙非丙型急性非乙非丙型慢性肝炎中的感染率最高;在TT病毒与乙型、丙型肝炎病毒混合感染的慢性肝炎中,TT病毒干涉乙型及丙型肝炎病毒造成的肝细胞损伤的可能性很小。     

Polymorphism of the TT virus and its frequency in Polish blood donors

OBJECTIVE: To determine, in Polish blood donors, the frequency of TT virus (TTV) using different primers and the sequence diversity of TTV genotypes. MATERIALS AND METHODS: Two-hundred blood donors were studied. TTV DNA was detected by the polymerase chain reaction (PCR) using primers for the coding (ORF1) and non-coding (NC) regions. Twenty isolates were genotyped by sequencing the ORF1 fragment. RESULTS: TTV DNA was detected in 78% of donors using NC primers and in 10% using ORF1 primers. The frequency of TTV DNA detection by NC primers was observed to increase with donor age, whereas the frequency of detection by ORF primers did not differ between various age-groups. The nucleotide sequence homology of Polish TTV isolates ranged from 59 to 99%. Three genotypes (1b, 2b and 2c) were identified. CONCLUSIONS: The frequency of TTV detection depends on the primers used for the PCR. Using the NC primers the virus is detected in the majority of donors, whereas the ORF1 primers strongly underestimate the prevalence of TTV. The frequency of TTV DNA increases with age. Polish TTV isolates are highly polymorphic and are classified as 1b, 2b and 2c.     

Clinical characteristics and distribution of genotypes of TT virus infection in a hepatitis C virus-hyperendemic township of a hepatitis B virus-endemic country (Taiwan)

BACKGROUND: The prevalence of TT virus (TTV) viremia, without definite clinical significance, has been reported to be higher among chronic hepatitis C patients. The status and clinical characteristics of TT virus (TTV) infection and distribution of TTV genotypes in a hepatitis C virus (HCV) hyperendemic township (Masago community) in a hepatitis B virus (HBV) endemic country (Taiwan) were investigated. METHODS: Sera from 100 Masago residents were tested for alanine aminotransferase (ALT) and markers of HBV, HCV and GB virus C/hepatitis G virus (GBV-C/HGV) and TTV-DNA. Sera of 250 blood donors as a control group were tested for TTV-DNA. Sera of Masago residents and blood donors with positive TTV-DNA were directly sequenced, and phylogenetic analyses were performed subsequently. RESULTS: The prevalences of TTV viremia in different age groups among individuals from Masago were significantly higher than that among blood donors. In regard to the subtypes of TTV, 23, seven, two, eight, one, six and one isolate were related to the genotypes 1a, 1b, 2a, 2b, 3, 4 and 5, respectively, from Masago and 21, 14, one, nine and three isolates were related to the genotypes 1a, 1b, 2a, 2b, and 4, respectively, from donors. No clinical or virological factor was associated with TTV viremia or TTV genotypes. CONCLUSIONS: TT Virus prevalence was higher among HCV hyperendemic township residents than blood donors with similar genotype distributions (genotype 1 was the most prevalent) in Taiwan. Neither TTV viremia nor a particular genotype was associated with HBV, HCV or GBV-C/HGV infection and abnormal ALT levels.