Detection of hepatitis C virus (HCV) RNA in paraffin embedded tissue sections of human liver of non-A, non-B hepatitis patients by in situ hybridization.

Localization of hepatitis C virus (HCV) RNA was investigated by non-radioactive in situ hybridization in human liver specimens of chronic non-A, non-B (NANB) hepatitis patients who were seropositive for antibodies to HCV (anti-HCV). For in situ hybridization, T-T dimerized synthetic oligodeoxynucleotide probes were used and DNAs hybridized in situ were detected immunohistochemically using specific antibodies against T-T dimer. The data demonstrates that HCV-RNA was localized in the cytoplasm of hepatocytes in human liver biopsies obtained from the patients with chronic NANB hepatitis seropositive for anti-HCV.     

Performance of an enzyme-linked immunosorbent assay system for antibodies to hepatitis C virus with two new antigens (c11/c7).

We developed an enzyme-linked immunosorbent assay (ELISA) system for antibodies to the hepatitis C virus (HCV), using two new recombinant antigens (c11 and c7) derived from the HCV genome. The performance of this ELISA system (Imucheck HCV Ab) was examined. The CV values for both intra-assay precision and reproducibility of identifying HCV antibody in the panel sera ranged from 3.5% to 6.4%. The blood elements in serum and anticoagulants did not interfere in this ELISA system. The specificity of Imucheck HCV Ab to samples from patients with non-A, non-B (NANB)-type chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma was 93.7%, 93.5%, and 81.4%, respectively. These results are more sensitive than those obtained by the first-generation anti-HCV ELISA system. In the samples from patients with NANB-type acute hepatitis, Imucheck HCV Ab enabled detection of HCV antibodies at an early stage. This system increased the sensitivity for blood donor screening and for monitoring patients with acute hepatitis.     

Hepatitis C virus RNA in blood units with antibodies detectable by a second-generation passive hemagglutination assay,antibodies to synthetic core peptides or elevated transaminase levels

Blood units from 3383 donors were tested for antibodies to hepatitis C virus (HCV) by a second-generation passive hemagglutination assay (PHA-2) with detector cells coated with three recombinant HCV proteins. They were tested also for antibodies to synthetic HCV core peptides (anti-CP9 and anti-CP10) by enzyme immunoassays and for alanine aminotransferase (ALT). PHA-2 was reactive in 32 (0.9%) units, high-titered anti-CP9 and/or anti-CP10 was detected in 101 (3.0%), and elevated ALT levels (> 45IU/L) in 99 (2.9%). At least one of these markers were detected in 209 units (6.2%), and HCV RNA was tested for in 197 of them. HCV RNA was detected in 21 units including 19 initially reactive on PHA-2 of which 17 were repeatedly reactive, 19 with anti-CP9 and/or anti-CP10 in high titers, and 5 with elevated ALT levels. Two units were reactive only on PHA-2, while the other two had anti-CP9/anti-CP10 as the sole indicator of hepatitis C viremia. These results indicated that PHA-2 and anti-CP9/anti-CP10 would be complementary in detecting blood units with HCV RNA, and effective in further decreasing the risk of post-transfusion hepatitis C.     

丙型肝炎病毒感染的献血者10年追踪观察

目的　追踪观察发生HCV感染的献血者的疾病进程和健康状况。方法　从 1993年 10月～ 2 0 0 4年 2月 ,对 30名HCV感染的献血者定期抽取血液标本 ,观察所得到的 4 4 2份系列标本的血清ALT、抗 HCV和HCV RNA的动态变化 ,并进行HCV分型测定 ,重点分析了其中 10名自愿者肝组织的病理检查结果和血清ALT、抗 HCV与HCV RNA的动态变化。结果　① 30名HCV感染者系列标本中 ,ALT异常率 37.6 % (16 6 / 4 4 2 ) ;抗 HCV阳性率97.1% (42 9/ 4 4 2 ,ELISA法 ) ;HCV RNA阳性率 74 .9% (331/ 4 4 2 )。② 10名HCV感染的献血者 10年系列血清中抗 HCV大多数持续存在 (94 4 % ,15 1/ 16 0 ) ,其HCV RNA时有间隙性阴性 ,阳性率 6 7 5 % (10 8/ 16 0 )其血清ALT水平异常率 33 6 % (5 0 / 14 9)。③HCV分型测定 :HCVⅡ / 1b型占 85 % (2 2 / 2 6 ) ,Ⅲ / 2a型占 15 % (4/ 2 6 )。④ 10名HCV感染者肝组织检查显示均为轻度慢性肝炎。结论　肝组织的病理变化同血清抗 HCV、HCV RNA和ALT的异常有明显相关性。虽然HCV感染后 10年的疾病进程大多是缓慢的 ,但应采取适当治疗措施以控制慢性肝炎的发展。     

Prevalence of hepatitis C infection in a large urban hospital-based sample of individuals with spinal cord injury

OBJECTIVE: To examine the prevalence and clinical characteristics of hepatitis C infection in individuals with chronic spinal cord injury (SCI). DESIGN: Retrospective case survey. SETTING: Outpatient clinic devoted to SCI follow-up care located in a county-government rehabilitation center. PARTICIPANTS: A total of 531 unselected individuals with chronic SCI. INTERVENTIONS: Patients underwent routine annual physical examinations at the outpatient clinic, and were tested for hepatitis C antibodies, antibodies to hepatitis core antigen, alanine aminotransferase (ALT), and bilirubin. MAIN OUTCOME MEASURES: Prevalence of hepatitis C antibodies and liver test abnormalities. RESULTS: Seventeen percent of the cohort was anti-hepatitis C virus (HCV) reactive (HCV positive). The prevalence of HCV infection in those who sustained SCI before 1990 was 21% compared with 7% (10/147) of those who were injured from 1990 onward (, P=.0002). Period of injury (Wald, P=.0042) and age (Wald, P=.048) were the only significant factors for anti-HCV reactivity. Thirty percent of the HCV-positive individuals had abnormal ALT levels compared with only 10% of the HCV-negative individuals (, P<.0001). Individuals who were HCV positive were more likely to be hepatitis B core antigen-reactive compared with those who were HCV negative (31% vs 9%;, P<.0001). CONCLUSIONS: The prevalence of HCV infection among individuals with chronic SCI is significantly higher than the general population. The majority of those with SCI and HCV infection have normal liver tests. Clinicians should maintain a high index of suspicion for HCV infection, even in the absence of elevated aminotransferase activities.     

Frequency and significance of antibody to hepatitis C virus in severe corticosteroid-treated autoimmune chronic active hepatitis.

To determine the frequency and significance of antibody to hepatitis C virus (anti-HCV) in severe autoimmune chronic active hepatitis, we tested sera from 85 cortico-steroid-treated patients by an enzyme immunoassay. Seropositive patients were assessed for specific antibodies to hepatitis C virus-encoded antigens by recombinant immunoblot assay. The findings in patients with and without anti-HCV were contrasted, and the frequency of seropositivity was compared with that in patients who had other types of chronic liver disease and in normal adults. Only 5 of the 85 patients with autoimmune hepatitis (6%) were seropositive for anti-HCV, and only 2 of these patients were reactive by recombinant immunoblot assay. The frequency of seropositivity in autoimmune hepatitis was not significantly different from that in hepatitis B surface antigen-positive (9%) and cryptogenic (18%) disease, but it was significantly less than that in posttransfusion chronic active hepatitis (6% versus 75%; P less than 0.001). Two patients became seronegative after corticosteroid therapy; both had been nonreactive by recombinant immunoblot assay. Four of the seropositive patients entered remission during corticosteroid therapy, including three whose sera were nonreactive to virus-encoded antigens. We conclude that anti-HCV occurs infrequently in corticosteroid-treated severe autoimmune hepatitis and that antibodies detected by enzyme immunoassay may be nonreactive to hepatitis C virus-encoded antigens. Seropositive patients who are nonreactive by immunoblot assay may still respond to corticosteroid therapy and become seronegative during treatment.     

Prevalence of hepatitis G virus RNA and antibodies to HGV envelope protein (anti-E2) in patients with liver injury in Japan]

To assess the role of hepatitis G virus (HGV) in acute and chronic liver diseases, we investigated the prevalence of HGV RNA and antibodies to HGV envelope protein (anti-E2) among patients with liver diseases diagnosed in our hospital from 1992 to 1997. Among 24 patients with acute hepatitis (HAV: 13, HBV: 2, HCV: 3, CMV: 1, non A-C: 5), only 1 patient with non A-C hepatitis (4%) were positive for HGV RNA and none was positive for anti-E2. Among 461 patients with chronic liver diseases (alcohol: 27, HBV: 74, HCV: 297, HBV + HCV: 10, non B non C: 14, autoimmune and metabolic: 39), 40 patients (alcohol: 1, HBV: 3, HCV: 33, HBV + HCV: 3) were positive for HGV RNA(9%) and 48 patients were positive for anti-E2(17%). In the patients with positive for anti-E2, only 8% were positive for HGV RNA. 98% of HGV RNA positive patients were infected with HBV or HCV, and especially 82% were infected with HCV. In patients with non A-C hepatitis, none was positive for HGV RNA, so HGV seems not to have important role in liver diseases.     

对丙型肝炎病毒感染的10名献血者的追踪研究

目的：了解感染丙型肝炎病毒（ Ｈ Ｃ Ｖ）献血者的预后。方法：选择刚发生 Ｈ Ｃ Ｖ 感染的３０ 名献血者,进行定期多项指标的动态追踪观察,对其中１０ 名自愿者作了肝组织病理检查。结果：１０ 名 Ｈ Ｃ Ｖ 感染者４ 年的１０３ 份系列血清检测及肝组织检查获得以下结果：①血清各种 Ｈ Ｃ Ｖ 抗体大多持续存在,１０ 人中无 Ｈ Ｃ Ｖ 抗体完全转阴者;②１０ 名感染者中 ９ 名血清 Ｈ Ｃ Ｖ Ｒ Ｎ Ａ 持续阳性或间断阳性,仅 １ 名在感染后１ 年转阴;③血清 Ａ Ｌ Ｔ 水平常有波动,１０ 名感染者１０３ 份血清 Ａ Ｌ Ｔ水平异常者占 ３０％ ;④１０ 名 Ｈ Ｃ Ｖ 感染者肝活检均表现为轻度慢性肝炎,７ 人的肝组织中有６ 人查到 Ｈ Ｃ Ｖ Ｒ Ｎ Ａ。结论：发生 Ｈ Ｃ Ｖ 感染的献血者大多有向慢性肝炎发展的趋势,因此对 Ｈ Ｃ Ｖ 感染的献血者应当积极采取适当治疗措施,以控制其向慢性肝炎发展。     

Testing for hepatitis C

Over the past 10 years, 12,146 cases of hepatitis were diagnosed in the Virology Department of Vienna University. 30.3% were hepatitis A, 39.2% hepatitis B, 3.0% cytomegalovirus and 1.5% Epstein-Barr virus infections. The remaining 25.8% were diagnosed as non-A, non-B hepatitis (NANB). Therefrom, a sample of 167 sera from acute and 78 from chronic hepatitis NANB were tested for hepatitis C. 9.6% of the acute and 44.9% of the chronic cases were positive. We conclude from these data that about 12% of all hepatitis cases in Austria are caused by the hepatitis C virus. In addition, risk groups for hepatitis C were tested. The highest prevalence (80%) was found in drug addicts. Polytransfused (organ transplanted) patients had antibodies in 44.8% of cases. Of 78 dialysis patients, 7 were positive but nearly all positives came from one single dialysis unit, thus indicating a prevalence of 30% there.     

Infective, chronic carriers of Hepatitis C virus among blood donors with no history of clinical hepatitis

Summary. We investigated the infectivity of three hepatitis C virus antibody (anti-HCV) positive blood donors with either hepatitis B core antibodies (anti-HBc) (Nos 1 and 2) or raised alanine aminotransferase (ALT) (No. 3). The 57 recipients of blood products from these donors during the period 1971–1990 were identified and the living 23 were tested for anti-HCV. Among these, 11 out of 14 (78%) recipients from Nos 1 and 2, and 1 out of 9 (11 %) recipients from No. 3 were anti-HCV positive. The former donors had high titres of anti-C 100-3 and high rating scores in the HCV recombinant immunoblot assay (RIBA). They were evidently infective, chronic carriers of HCV but had no clinical signs or medical history of hepatitis. The latter donor had low titres of anti-C100-3 and a low RIBA rating score. She had clinical signs of chronic hepatitis and persistently elevated ALT, but only one of her recipients was anti-HCV positive.     

High frequency of HCV infection in individuals with isolated antibody to hepatitis B core antigen

Although isolated antibody to hepatitis B core antigen (anti-HBc) is frequently nonspecific or may be the only serological marker of past self-limiting hepatitis B, where antibodies against the surface antigen have disappeared, isolated anti-HBc seropositivity is frequently associated with chronic hepatitis B in HIV- and HCV-infected individuals. Of 5,520 samples that tested positive for anti-HBc (IMx and AxSYM CORE, Abbott, Delkenheim, Germany) at the Institute of Virology, University Clinic Frankfurt during the time interval from January 1994 to February 1996, 643 (11.6%) were isolated anti-HBc-reactive in the IMx and AxSYM CORE assays (inhibition values >90%). There was a statistically significant association between isolated anti-HBc seropositivity and HCV and HIV/HCV coinfection (p < 0.05). A total of 190 samples were available for further testing. Six (3.2%) of 190 isolated anti-HBc-positive samples were considered false-positive since they were only positive in the AxSYM or IMx CORE assay and a linear decrease of the measured signal could not be observed in dilution series. Of 184 serum samples tested with nested PCR using primers of the S genome region, only 6 (3.3%) were HBV DNA-positive. Anti-HBc-IgM antibody could be detected in 3 (1.6 %) of the tested samples using the IMx CORE-M. With the more sensitive VIDAS HBc IgM specific IgM antibody was detected in 15 (8.5%) of 177 samples at concentrations ranging from 10 to >200 Paul Ehrlich Institute U/ml. HIV or HCV coinfection was present in 28.1% and 37.5% of isolated anti-HBc-positive individuals, respectively. We conclude from our observations that only a limited proportion of anti-HBc-isolated individuals are potentially infectious, however anti-HBc-IgM which is detectable in any form of liver disease associated with HBV infection was present in more than 8% of the individuals. Of isolated anti-HBc-positive sera 37% were positive for anti-HCV, suggesting that anti-HCV antibody testing should be performed in isolated anti-HBc-positive individuals.     

两种荧光定量法检测HCV RNA结果的比较

目的建立双探针荧光定量HCV RNA检测方法,分析其定量检测HCV RNA的灵敏度和特异性以及HCV RNA含量与患者病情的相关性。方法选取100例抗HCV抗体阳性患者,包括慢性肝炎患者45例、肝硬化患者30例、肝癌患者25例的样本,用双探针荧光定量法和两种商品荧光定量试剂方法进行HCV RNA检测,另观察近期196例抗HCV抗体阳性和48例抗HCV抗体阴性样本荧光定量PCR HCV RNA含量与肝功能的相关关系。结果荧光定量双探针法阳性率为93％（93／100）;两种商品荧光定量试剂方法阳性率分别为84％（84／100）和79％（79／100）,经卡方检验差异有统计学意义（P〈0．05）。196例中抗HCV与HCV RNA阳性一致率为57．1％（116／196）,其中伴肝功能改变49．0％（96／196）。HCV RNA含量与AST和ALT异常程度无显著相关性（P〉0．05）,相关系数分别为0．4293及0．3899。HCV RNA高拷贝数患者肝功能异常率为85．2％（69／81）,低拷贝数者肝功能异常率为40．9％（47／115）,经卡方检验,X^2=38．63,差异有统计学意义（P〈0．001）。结论双探针荧光定量提高了HCV RNA检测的灵敏度和特异性,HCV RNA含量与患者肝功能损伤程度无明显相关性。     

Performance of a conventional enzyme immunoassay for hepatitis C virus core antigen in the early phases of hepatitis C infection

There are periods within the early phase of hepatitis C virus (HCV) infection in which the anti-HCV antibody test is unable to confirm HCV viremia. To reduce the risk of transmitting HCV through transfusions, we developed a simple and highly sensitive enzyme immunoassay (EIA) which detects the core antigen of HCV (HCVcAg). This assay employed a conventional colorimetric EIA system, and was based on a two-step sandwich assay, using a 96- well microplate. The reproducibility of the results was very high. When the cutoff values were set to 30 fmol of recombinant HCVcAg/L, as determined by the distribution of healthy subject sera (n=223), 99.6% of healthy subject sera and 100% of hepatitis B patient sera (n=50) were negative for HCVcAg. The clinical performance of this EIA was examined using 14 commercially available seroconversion panels. In every panel, HCVcAg could be detected at points preceding the seroconversion of anti-HCV antibodies. The points at which HCVcAg was detected were the same as those at which it was detected by an AMPLICOR HCV Monitor test. The EIA's window period for detecting the HCVcAg in all panels was on average 26 days shorter than that of the anti-HCV antibody test. In three panels where the first sample is negative for HCV RNA, the window period was shortened 50 days by this EIA for HCVcAg. There was a positive correlation between the concentration of HCVcAg and HCV RNA in anti-HCV antibody negative specimens. This assay was simpler to perform than assays based on gene amplification technology for the detection of HCV RNA, and the window period was shortened to that of the AMPLICOR HCV Monitor test. Thus, the EIA for HCVcAg would be useful in screening seroconverting donors and could reduce the residual risk of secondary HCV infections through transfusions.     

Immunochemical and biomolecular studies of circulating immune complexes isolated from patients with acute and chronic hepatitis C virus infection

Circulating immune complexes (ICs) were isolated by affinity chromatography and sucrose density gradient fractionation during acute and chronic hepatitis C virus (HCV) infection. Immunochemical and biomolecular studies showed that they basically consist of the virus component, IgG with specific anti-HCV activity and IgM bearing 17.109 epitope (IgM 17.109), an antigenic determinant common to rheumatoid factors (RFs) with WA cross-idiotype (XId). An antigen-specific IC assay was used to demonstrate IgG anti-HCV/IgM 17.109 ICs (IgG-IgM ICs) in five out of the five patients with acute and in 8 out of the 10 patients with chronic hepatitis C who mounted an IgG anti-HCV immune response. They were not detected in patients with no IgG anti-HCV response. IgG–IgM ICs appeared in step with IgG anti-HCV seroconversion and remained detectable for a long period irrespective of clinical outcome, in that they were demonstrated over a 4-year follow-up of patients with chronic hepatitis C. Their presence was unrelated to the severity and progression of liver histology. Despite similar serum levels of IgM 17.109 XId, antigen-specific IgG-IgM ICs were not found in acute and chronic hepatitis B or in acute hepatitis A. Thus, these ICs appear to be uniquely associated with HCV infection, supporting the view that IgM 17.109 XId derive from an antigen-driven response strictly related to the involved antigen. Even although they have no apparent effects on the progression of HCV-related liver disease, their presence may help to explain the immunological abnormalities and extrahepatic disorders observed in HCV infection.     

Seroprevalence of hepatitis C virus (HCV) infection among blood donors: a hospital-based study.

Insofar as chronic hepatitis C virus (HCV) infection in many individuals is asymptomatic, and as the prevalence of antibodies to hepatitis C virus (anti-HCV) among blood donors in Lebanon is scarce, this study addressed the prevalence of anti-HCV in 5,115 blood donors. Data obtained were compared to other world regions. Of the blood donors screened, 57 were initially tested positive or doubtful for anti-HCV Ab. Subsequent testing by two-third generation enzyme immunoassays confirmed that, of the 57 initially tested positive/doubtful, only 18 were positive for anti-HCV giving a prevalence rate of 0.4%. While there was no difference in HCV prevalence with respect to age or gender, a higher rate was seen in non-Lebanese compared to Lebanese subjects (3.4% vs 0.3%, P < 0.001). These results demonstrate a low prevalence of HCV infection among Lebanese blood donors, which was comparable to those established for western countries.     

丙型肝炎病毒RNA含量与抗-HCV及ALT的关系

目的 了解丙型肝炎病毒核糖核酸(HCV RNA)与丙型肝炎病毒抗体(抗-HCV)及丙氨酸氨基转移酶(ALT)的关系.方法 144例在本院住院和门诊同时检测HCV RNA、抗-HCV及ALT的患者,用荧光定量逆转录聚合酶链反应(FQ-RT-PCR)法检测标本中的HCV RNA,同时采用酶联免疫吸附试验(ELISA)法检测抗-HCV,用全自动生化检测仪测定ALT水平.结果 144例标本中HCV RNA高于上限值34例(23.6%),抗-HCV阳性44例(30.6%),二者有很好的相关性.有46例(31.8%)存在ALT异常,而ALT异常率与HCV RNA含量有一定的比例关系.结论 HCV RNA含量与抗-HCV同时检测可提高临床对丙型肝炎的诊断.HCV RNA是反映HCV复制的可靠指标,结合ALT结果可帮助临床了解HCV在体内的复制状况及肝脏的炎性反应状态.FQ-RT-PCR法检测HCV RNA具有非常好的临床应用价值.     

Lichen planus and hepatitis C virus in the Northern Kyushu region of Japan

Abstract. Oral lichen planus (OLP) is a common oral disorder that manifests a mucosal reaction to a variety of aetiological factors, including liver disorder. This study investigated the relationship between OLP and hepatitis C virus (HCV) infection by studying the prevalence of hepatitis B and C virus infection or liver disease in 45 patients with OLP in the Northern Kyushu region of Japan where the prevalence of HCV infection is the highest in the country. Serum hepatitis B virus surface antigen (HBsAg) was positive in only four patients. Serum anti-HCV or serum HCV RNA was positive in 28 (62%) and 27 (60%) of 45 OLP patients, respectively. The majority (35 of 45, 78%) of OLP patients suffered from liver disease, including chronic hepatitis C (22/45, 49%), HCV-related liver cirrhosis (two), and HCV-related hepatocellular carcinoma (two). These results suggest that HCV is a major cause of OLP.     

Lack of correlation between sensitivity characteristics of the tests for hepatitis C virus antibodies estimated with serially diluted and natural low-reactive control specimens

Sensitivity characteristics of seven commercial ELISA test systems for the detection of antibodies to hepatitis C virus were assessed using control panels consisting of: (i) serial dilutions of pooled sera highly reactive for anti-HCV; (ii) serial dilutions of RIBA 3.0 HCV SIA positive control; and (iii) natural (non-diluted, non-spiked) sera low-reactive for anti-HCV. "Dilutional sensitivity" values estimated with these two kinds of highly reactive samples did not coincide and were not found to correlate with the proportion of natural low-reactive specimens detected by each test. Thus, laboratories assessing sensitivity of anti-HCV ELISAs should take into consideration the nature and properties of the control material used. Natural low-reactive control specimens are preferable because they adequately reflect the real serological picture of early stage of HCV infection.