安徽省不同人群及各型肝炎患者庚型肝炎病毒感染检测分析

应用酶联免疫试验（ＥＩＡ）和逆转录套式聚合酶链反应（ＲＴ－ｎＰＣＲ）对１００ 例一般人群、３８５例献血员、５４ 例血液透析患者、７２ 例乙型肝炎、４１ 例丙型肝炎２７ 例非甲－戊型肝炎患者进行检测。结果抗－ＨＧＶ 阳性率分别为２．００％ 、７．５３％ 、２７．７８％ 、１８．０６％ 、１９．５１％ 和１４．８１％ ;抗－ＨＧＶ 阳性者中ＨＧＶＲＮＡ 阳性率分别为１００．００％ 、６２．０７％ 、６６．６７％ 、６９．２３％ 、７５．００％ 和 １００．００％ ,提示本地区不同人群存在ＨＧＶ 感染。献血员、血透患者、乙型肝炎、丙型肝炎、非甲－戊型肝炎患者的ＨＧＶ 感染率显著高于一般人群,提示献血员,血透患者及ＨＢＶ、ＨＣＶ 感染者是ＨＧＶ 感染的高危人群。ＨＧＶ 常与ＨＢＶ 或ＨＣＶ 重叠／联合感染,也可单独感染。抗－ＨＧＶ 阳性者中ＨＧＶ ＲＮＡ 阳性率为８３．８２％ ,提示抗－ＨＧＶＥＩＡ 可用于ＨＧＶ 感染的检测。ＡＬＴ 正常和异常献血员中抗－ＨＧＶ 阳性率无显著性差异。     

肝炎病毒与EB病毒重叠感染

为探讨肝炎病毒（ＨＶ）与ＥＢ病毒（ＥＢＶ）重叠感染的状况和后果,我们用免疫酶法对１５４例各型病毒性肝炎患者作了ＥＢＶＩｇＡ抗体检测。结果发现,急性肝炎、慢性轻度肝炎、慢性中度肝炎、肝炎肝硬化、慢性重型肝炎和原发性肝癌ＶＧＡ－ＩｇＡ抗体的阳性率分别为２４．０％、３０．０％、５３．３％、６３．３％、４０．０％和７２．７％,与健康人（５．３％）比较,有非常显著升高（Ｐ＜０．０１）;原发性肝癌又较急性肝炎和慢性轻度肝炎高,并有非常显著意义差异（Ｐ＜０．０１）。ＨＢＶ和ＨＡＶ＋ＨＢＶ感染者比较,前者又较后者低（Ｐ＜０．０１）。重叠感染者的临床表现均为“肝炎型”,未见咽炎、腺热、胃肠、肺炎、肾炎、神经等类型。重叠感染者的ＣＤ＋３及ＣＤ＋４Ｔ细胞下降,ＣＤ＋８Ｔ细胞及ＩｇＧ,ＩｇＭ升高,与健康人比较差异非常显著意义（Ｐ＜０．０１）。结果提示：ＨＶ感染,不仅因免疫失调易感ＥＢＶ,又可因重叠感染而进一步使免疫功能失调;对病毒性肝炎的处理应强调免疫调节治疗。     

丙型肝炎病毒颗粒研究进展

本文综述了近年来对丙型肝炎病毒（ＨＣＶ）颗粒的研究进展。在ＨＣＶ感染引起的肝炎组织标本、血液标本及培养细胞中,普通透射电镜下发现有病毒颗粒的存在。核内颗粒直径为２０￣２７ｎｍ;胞质内颗粒的大小报道各异,其结构包括核心和外壳两部分,核心平均直径４０ｎｍ。这种颗粒的特性尚未被阐明。近年在ＨＣＶ感染的培养细胞及黑猩猩ＨＣＶ肝炎模型中免疫电镜检查证明这种颗粒有病毒抗原性,但在人们ＨＣＶ肝炎肝组织中这种颗料     

丙型肝炎病毒RNA和丙型肝炎病毒抗体在慢性丙型肝炎干扰素治…

干扰素是慢性丙型肝炎抗病毒治疗的首选药物。近年来,丙型肝炎病毒（ＨＣＶ）ＲＮＡ和抗－ＨＣＶ检测方法的建立为干扰素疗效监测提供了新的手段。本文就这方面的进展作简要综述。     

两例丙型肝炎病毒(HCV)和乙型肝炎病毒(HBV)重叠感染者的HCV核心区cDNA序列分析

从临床肝病患者中选择两例ＨＣＶ和ＨＢＶ重叠感染者ＨＳＱ和ＳＺＨ,他们血清中的生化指标丙氨酸转氨酶（ＡＬＴ）持续异常,肝活检病理示有严重的肝损伤。在ＡＬＴ异常期,血清学检测结果为ＨＢｓＡｇ、ＨＢｅＡｇ阳性,抗ＨＣＶＩｇＧ（包括Ｃ２２、Ｃ３３ｃ）阴性,但套式ＰＣＲ检测ＨＣＶＲＮＡ阳性,核心区ｃＤＮＡ序列分析发现该区有１个密码子（ＧＧＣｎｔ３８５—３８７）缺失,对应缺失的氨基酸是甘氨酸（ＧＬＹ）,从血清学检测和序列分析结果推测,在ＨＣＶ和ＨＢＶ重叠感染中,ＨＢＶ和ＨＣＶ均可处于持续复制状态,抗ＨＣＶＩｇＧ抗体阴性可能是ＨＣＶ的多蛋白前体翻译和病毒颗粒装配受到ＨＢＶ干扰的结果。     

福州地区庚型肝炎病毒重叠感染

为研究庚型肝炎病毒在福州地区的重叠感染,采用ＥＬＩＳＡ法检测本院住院的２８６例病毒性肝炎（ＨＶ）患者和５００名供血员的抗－ＨＧＶ。结果表明,甲、乙、丙、戊型肝炎患者和供血员的抗－ＨＧＶ检出率分别为２．０％、２．２％、４．０％、１０．０％和０．２％。急性肝炎、慢性肝炎、慢性重型肝炎、肝硬化、原发性肝癌和抗－ＨＣＶ阳性供血员的检出率分别为７．９％、４．３％、３３．３％、０％、７．１％和６．３％,慢性重型肝炎检出率较慢性肝炎显著升高（Ｐ＜０．０５）。各型肝炎患者和供血员均存在庚型肝炎病毒重叠感染,以慢性重型肝炎为著。     

丙型肝炎病毒与自身免疫性肝炎

丙型肝炎病毒（ＨＣＶ）特异检测方法建立后发现自身免疫性肝炎（ＡＩＨ）抗－ＨＣＶ检出率高,有人提出ＨＣＶ感染有可能为ＡＩＨ的启动因素之一,尤为Ⅱ型ＡＩＨ。区别抗－ＨＣＶ阳性的肝炎为丙型肝炎还是ＡＩＨ十分重要,因两者的现行治疗措施完全不同。     

丙型肝炎病毒感染标志检测临床意义的研究现状

随着丙型肝炎病毒分子生物学研究的深入，相继建立了多种ＨＣＶ感染标志的检测方法。本文就ＩｇＧ抗－ＨＣＶ，ＨＣＶＲＮＡ和ＩｇＭ抗－ＨＣＶ检测在丙型肝炎诊断，预后估计及抗病毒药物疗效评价方面的意义作简要综述。     

SABC法和胶体金标免疫电镜观察体外感染细胞中HCV—NS5的表达

寻找敏感的丙型肝炎病毒 (ＨＣＶ)体外培养系统 ,对于研究ＨＣＶ的病毒体特征、致病机理、抗病毒治疗和疫苗研制等方面有着重要的意义。我们曾在体外感染的人Ｔ淋巴细胞中发现ＨＣＶ的正、负链[1] ,随后对于ＨＣＶ在体外感染细胞中的抗原表达情况又做了进一步研究 ,现报道结果如下。1　材料和方法1.1　ＨＣＶＲＮＡ阳性接种物选择ＨＣＶＲＮＡ水平为 2× 10 4 拷贝 /ｍＬ(荧光定量法。试剂盒由美国Ｂｉｏｔｒｏｎｉｃｓ公司提供 )的血清 ,该血清检查甲、乙、丁、戊型肝炎标志为阴性 ,以此作为接种物 ,同时 ,选用正常人的血清作阴性对照。1.2…     

献血员血清抗—HCV和HCV RNA检测

本文对宜昌市的２９９９名献血员,５９４名自然人群的血清进行了抗－ＨＣＶ、ＨＣＶＲＮＡ检测,结果表明,献血员中抗－ＨＣＶ阳性２３例,阳性率为０．７７％。其中宜昌、巴东分别为０．７３％和０．７６％,沙市为０．８３％。地区、性别、城乡之间抗－ＨＣＶ阳性率无显著性差异（ｐ＞０．０５）。参照以往文献报导,此阳性率比国内报导的低,与国际的相一致。５９４名自然人群抗－ＨＣＶ结果均为阴性,献血员与自然人群之间抗－ＨＣＶ阳性率差异显著（ｐ＜０．０５）。２０例抗－ＨＣＶ阳性者中检出ＨＣＶ　ＲＮＡ阳性６例,占３０％（６／２０）,占受检献血员的０．２０％（６／２９９９）,比武汉（０．８０％）低４倍,地区之间无显著差异（ｐ＞０．０５）。由此初步确认：宜昌市献血员中丙型病毒性肝炎感染低于国内某些大中城市、宜昌市（含七县二市）巴东县、沙市市是丙型病毒性肝炎低感染区。但存在无症状病毒血症携带者传染源。     

丙型肝炎患者血浆和外周血单个核细胞同步检测HCV-RNA和HGV-RNA

To understand the incidence of hepatitis C and hepatitis G coinfection, and positive rate of HCV-RNA or HGV-RNA in plasma and PBMC, HCV-RNA and HGV-RNA in plasma and in peripheral blood mononuclear cells(PBMC) of the 40 patients were amplified with RT-PCR. There were 6 and 8 HGV-RNA positive cases in plasma and PBMC, respectively. And there were 5 HGV-RNA positive cases both in plasma and PBMC.At the same time, there were 5,6,3 both HCV-RNA and HGV-RNA positive cases in plasma, in PBMC and in both plasma and PBMC,respectively. It accounted for 13%,15% and 8%, respectively. The HCV-RNA and HGV-RNA positive incidence of PBMC was higher than that of plasma. The incidence of HCV-RNA and HGV-RNA coinfection was similar to European, American and Japanese. The synchronous detection has an important signficance in avoiding leaked diagnosis.     

乙型肝炎病人重叠感染丙型肝炎的评价

应用ＥＬＩＳＡ和ＰＣＲ法检测５０２例乙肝病人血清,４０１例ＨＢｓＡｇ阳性血清中,有１１４例（２８．４％）抗－ＨＣＶ和ＨＣＶＲＮＡ双项阳性,２５例（６．２％）ＨＣＶＲＮＡ单项阳性;２１例（５．２％）抗－ＨＣＶ单项阳性。将ＨＢｓＡｇ乙肝病人分成ＨＢＶＤＮＡ,ＨＢｅＡｇ阳性组和ＨＢＶＤＮＡ,ＨＢｅＡｇ阴性组。前者抗－ＨＣＶ阳性率为１１．６％～２０．５％,ＨＣＶＲＮＡ阳性率为１６．２％～２０．５％。后者抗－ＨＣＶ阳性率为２０．２％～５５．６％,ＨＣＶＲＮＡ阳性率为２３％～６０．３％。结果说明长期携带ＨＢＶ者和慢性乙肝病人均可重叠ＨＣＶ感染。ＨＢＶＤＮＡ阳性组抗－ＨＣＶ和ＨＣＶＲＮＡ阳性率明显高于ＨＢＶＤＮＡ阳性组     

RT套式PCR检测血浆HCV RNA及与抗HCV检测的比较

应用微量血清热变性法提取核酸,逆转录套式聚合酶链反应(RT-nest PCR)检测血浆HCV RNA,并与抗HCV ELISA检测结果比较,对HCV RNA阳性标本进行HGV RNA的筛查.结果在32例抗HCV阳性和20例抗HCV阴性血浆中,HCV RNA分别检出18例和2例,总符合率为70%,20例HCV RNA阳性者中有2例合并感染HBV,1例合并感染HGV.证明血浆样本中抗HCV与HCV RNA间存在很大的相关性.     

Detection of HCV and GBV-CHGV RNA in peripheral blood mononuclear cells of patients with chronic type C hepatitis

Hepatitis C virus (HCV) and hepatitis G virus (HGV) viraemia were investigated by RT-PCR protocols in peripheral blood mononuclear cells (PBMC) of 22 patients with chronic type C hepatitis. Samplings were at basal and 4-8 months after a 12 month period of treatment with interferon-alpha. A plus strand of HCV in PBMC was detected in 8 of 21 patients (38%) (p <0.05; chi2 test) with a lack of response to therapy; a minus strand was detected in 10% of chronic type C hepatitis and 25% of the patients harboured HCV RNA in PBMC. The association with a response was nearly significant (p <0.1; chi2 test). GBV-C/HGV RNA was detected in the serum of 9 of 21 (43%) patients and in PBMC of 20% of the patients viraemic. Genomic sequences of GBVC/HGV in PBMC were found, but further investigation is needed to assess the findings reported for HCV.     

Lack of evidence for hepatitis G virus replication in the livers of patients coinfected with hepatitis C and G viruses.

The pathogenic implications of hepatitis G virus (HGV) infection are still unclear. We searched for the presence of HGV RNA and HCV RNA sequences in liver and serum samples from 10 patients with chronic liver disease, 9 of whom were coinfected with HCV. All livers were negative for the presence of the HGV RNA minus strand and only six were positive for the presence of the positive strand, albeit at low levels. In striking contrast, the HCV RNA positive strand was detectable in the liver samples from all nine HCV-positive patients in titers ranging from 10(2) to 10(8) genomic eq/microg of RNA, and the negative HCV RNA strand was present in all but two of these patients. However, the positive-strand RNA titers in serum for the two viruses had similar ranges. These findings imply that the liver is not the primary replication site for HGV, at least in the population of HCV/HGV-coinfected patients. Absence of replication in liver tissue may explain the reported lack of influence of HGV coinfection on the course of chronic hepatitis C.     

Hepatitis G virus (GBV-C) infection among Brazilian patients with chronic liver disease and blood donors

Background: The recently discovered hepatitis G virus (HGV) belongs, as hepatitis C virus (HCV), to the Flaviviridae family. HGV has been isolated from the serum of patients with non A-E hepatitis. However, the association of HGV with hepatitis is uncertain.Objective: To determine the HGV prevalence in blood donors and in patients with liver disease and to evaluate a possible correlation between HGV infection and liver disease.Study design: Sera from a total of 113 consecutive patients with chronic liver disease were submitted to a series of liver enzymes and function tests and analyzed for the presence of HBsAg, anti-HBs, anti-HBc, anti-HCV, HCV RNA and HGV RNA. Prevalence of HGV RNA was determined in a group of 87 blood donors.Results: Nine (10%) sera from blood donors and 15 (13%) sera from patients with chronic liver disease were HGV RNA positive. Some 28 (25%) patients were HCV RNA positive, with genotypes 1a, 1b and 3 present in 10, 12 and 5 patients, respectively. A total of 20 (18%) patients were HBsAg carriers. Five (4%) patients were double infected (one with HBV+HCV, one with HBV+HGV and three with HCV+HGV).Conclusion: The proportion (10%) of HGV-infected blood donors was very high when compared with other countries. The results did not allow to establish HGV as an etiologic agent for chronic liver disease. The parenteral route was the presumed means of HGV transmission for only one-third of the patients.     

Co-infections with hepatitis G and TT virus in patients with chronic hepatitis C in Hungary

The significance of co-infections with novel hepatitis viruses Hepatitis G (GBV-C, HGV) and TT virus (TTV) in chronic hepatitis C is not clear. We determined the prevalence of HGV RNA and TTV DNA in chronic hepatitis C patients and in asymptomatic hepatitis C virus (HCV) carriers, and assessed the influence of these agents on the course of HCV infection. Seventy-seven patients with chronic hepatitis C--50 of them treated with interferon (IFN)--and 33 HCV carriers with normal alanine aminotransferase have been investigated. Previous HBV infection was detected by testing serum HBsAg and aHBc. HGV RNA and TTV DNA were detected by PCR. In the healthy population, the prevalence of anti-HCV was 0.3%, HGV RNA 8.0% and TTV DNA 18.5%. In chronic hepatitis C HGV RNA occurred in 9.09% and TTV DNA in 40.25% of cases. In IFN-treated patients with sustained remission, the frequency of TTV was 20% vs. 45.7% found in non-responders. Among asymptomatic HCV-carriers, the prevalence of HGV RNA was 9.09% and TTV DNA 75.7%. Neither HGV RNA nor TTV DNA had apparent effect on the HCV infection. TTV was detected with the lowest frequency in persons with sustained remission due to IFN, suggesting antiviral effect of IFN on TTV.     

Persistence of Hepatitis C Virus Traces after Spontaneous Resolution of Hepatitis C

Hepatitis C virus (HCV) frequently causes chronic hepatitis, while spontaneous recovery from infection is infrequent. Persistence of HCV after self-limited (spontaneous) resolution of hepatitis C was rarely investigated. The current study aimed to assess incidence and robustness of HCV persistence after self-resolved hepatitis C in individuals with normal liver enzymes and undetectable virus by conventional tests. Applying high sensitivity HCV RNA detection approaches, we analyzed plasma and peripheral blood mononuclear cells (PBMC) from individuals with previous hepatitis C infection. Parallel plasma and PBMC from 24 such non-viraemic individuals followed for 0.3–14.4 (mean 6.4) years were examined. Additional samples from 9 of them were obtained 4.5–7.2 (mean 5.9) years later. RNA was extracted from 250 μl plasma and, if HCV negative, from ~5 ml after ultracentrifugation, and from ex vivo stimulated PBMC. PBMC with evidence of HCV replication from 4 individuals were treated with HCV protease inhibitor, telaprevir. HCV RNA was detected in 14/24 (58.3%) plasma and 11/23 (47.8%) PBMC obtained during the first collection. HCV RNA replicative strand was evident in 7/11 (63.6%) PBMC. Overall, 17/24 (70.8%) individuals carried HCV RNA at mean follow-up of 5.9 years. Samples collected 4.5–7.2 years later revealed HCV in 4/9 (44.4%) plasma and 5/9 (55.5%) PBMC, while 4 (80%) of these 5 PBMC demonstrated virus replicative strand. Overall, 6/9 (66.7%) individuals remained viraemic for up to 20.7 (mean 12.7) years. Telaprevir entirely eliminated HCV replication in the PBMC examined. In conclusion, our results indicate that HCV can persist long after spontaneous resolution of hepatitis C at levels undetectable by current testing. An apparently effective host immune response curtailing hepatitis appears insufficient to completely eliminate the virus. The long-term morbidity of asymptomatic HCV carriage should be examined even in individuals who achieve undetectable HCV by standard testing and their need for treatment should be assessed.     

Prevalence of GB virus C/hepatitis G virus in Hungary

In 1995 a new flavivirus, GB virus C/hepatitis G virus (GBV-C/HGV), was discovered. The aim of this study was to determine the prevalence of the virus in healthy persons and hepatitis patients in Hungary. The sera of 408 healthy persons older than 60 years were tested for the presence of GBV-C/HGV antibodies, and 113 were positive (28%). Eight of the 71 healthy persons younger than 60 years and twenty of the 51 sera (39%) taken from patients suffering from hepatitis of unknown origin proved to be positive for GBV-C/HGV antibodies. Ten of the 124 sera (8%) of healthy persons and 36 of the 247 sera (14.6%) of hepatitis patients proved to be positive for GBV-C/HGV RNA. Eleven PCR products were sequenced, and the sequences were found to be different from each other and from the previously published ones. However, three sequences taken from the same patient at different times were identical. These results show that GBV-C/HGV is present in Hungary and cannot be considered rare.