A comparison of hepatitis C virus (HCV) RNA and – antibody as markers of infection and predictors of infectivity

Background: The diagnosis of hepatitis C virus (HCV) infection currently relies on the detection of antibody to HCV (anti-HCV). However, anti-HCV positivity may indicate past infection, current infection or possibly non-specific reactivity. For confirmation of current infection the virus needs to be assayed directly and this is possible by the polymerase chain reaction (PCR). Aims: The aims were to compare HCV RNA and anti-HCV as markers of infection in two groups of individuals: (i) a heterogeneous group with suspected HCV infection and (ii) a small group of blood and bone marrow donors, and their respective recipients. Methods: Serum samples were tested for alanine aminotransferase (ALT) as part of a liver function screen, for anti-HCV by ELISAII, and HCV RNA was detected by PCR. Single round and nested PCR was performed using primers designed from the sequence of the 5′-untranslated region of the HCV genome. Results: Of the 36 subjects in the heterogeneous group, 19/22 anti-HCV-positive patients with chronic non-A non-B hepatitis (NANBH) were viraemic, and the majority (17/19) demonstrated elevated ALT. However, HCV RNA was undetected in seven anti-HCV-positive patients, four of whom suffered autoimmune hepatitis Type I and three were low risk blood donors. Of the remaining subjects (seven/36) who were anti-HCV-negative, three/seven were HCV-RNA-positive and included two with acute post-transfusion (PT) NANBH and a recent needlestick victim who contracted HCV. In the second group, four individuals (donors), including a mother with a history of drug use, were implicated in transmission to three recipients. ALT levels were normal in all donors but raised in two of the recipients. PCR determined which of two anti-HCV-negative blood donors was infectious, confirmed transmission between a bone marrow donor and recipient, and indicated that anti-HCV detected in a newborn child represented passive transfer of antibody. Conclusions: Anti-HCV detected by ELISA II is a useful marker of chronic HCV infection, particularly in association with raised ALT. However, HCV RNA is a superior marker of acute HCV infection, a more reliable predictor of infectivity and is more specific.     

Risk of hepatitis C virus infection among household contacts of Saudi patients with chronic liver disease

SUMMARY. To evaluate the intrafamilial transmission of heptitis C virus and related risk factors among the Saudi population, two groups were investigated: 120 index patients with chronic liver disease and their 127 family contacts, and 220 blood donors who were anti-HCV-positive but with no chronic liver disease and their 91 family contacts. After a questionnaire on the risk factors for parenteral exposure, blood samples were obtained and tested for liver biochemistry and antibody to HCV (anti-HCV) by a third-generation enzyme immunoassay (UBI HCV HA4.0). Only two spouses of 20 index patients were anti-HCV-positive while the remaining 125 family contacts were anti-HCV-negative. None of the 91 family contacts of the 20 anti-HCV-positive blood donors was anti-HCV-positive. The two spouses were wives of index patients but had a history of blood transfusion on at least two different occasions. Our results clearly indicate the intrafamilial transmission of HCV is not the route of transmission of HCV among Saudis and our results argue against sexual transmission of hepatitis C virus despite a relatively long duration of marriage.     

Epidemiology,serological markers,and hepatic disease of anti-HCV ELISA-2-positive blood donors

The epidemiology associated with hepatitis C virus (HCV) infection, serologic reactivity, and hepatic disease related to anti-HCV-positive donors of Granada were researched. From 1990 through 1993, medical and epidemiological information and anti-HCV and HCV RNA testing were evaluated in 46,741 blood donors. Serum samples were obtained for anti-HCV ELISA and RIBA and HCV RNA determination. A liver biopsy was conducted in all anti-HCV positives by confirmatory second-generation RIBA to analyze the hepatic lesion and the presence of HCV RNA. The anti-HCV prevalence was 1.12%. A total of 228 anti-HCV second-generation ELISA positive blood donors were analyzed. Intrafamiliar transmission rate was 1.7%. Transfusion and intravenous drug abuse (IVDA) antecedents were associated with a higher risk of seroconversion. A RIBA-positive result was related to high second- and third-generation ELISA ratios (90%), HCV RNA positivity (89%), and elevated alanine aminotransferase (ALT) levels (88%). Approximately 50% of donors with normal ALT levels had high ELISA ratios and second-generation RIBA and HCV RNA positive results. Of the second-generation RIBA indeterminate results, 42% and 82% of the c22 and 33% and 100% of the c100 reactivities were third-generation RIBA and HCV RNA positive, respectively. Liver biopsy was conducted in 85 donors, 74% of whom had a chronic hepatitis and 83% had detectable HCV RNA levels. Chronic hepatitis was diagnosed in 88% vs 43% of donors with elevated and normal alanine aminotransferase levels, respectively. ELISA and confirmatory HCV RNA determinations should be routinely employed in donor screening. A liver biopsy should be conducted in patients with elevated ALT levels and normal ALT levels when viremic.     

Antibody to Hepatitis C Is Common among Patients with Alcoholic Liver Disease with and without Risk Factors

Thirty-seven patients with clinically suspected alcoholic liver disease were retrospectively studied for the prevalence of antibody to hepatitis C virus (HCV) by enzyme-linked immunosorbent assay (ELISA) and immunoblot assay. Twenty-four had biopsy-proven cirrhosis. Nineteen had identifiable risk factors for non-A, non-B viral hepatitis, and 18 did not. Five of 19 high-risk (26%) and 6 of 18 low-risk (33%) patients had positive antibody, compared with two of 179 healthy blood donors (p less than 0.01 for either group of alcoholics compared with blood donors). Nine of 11 ELISA-positive patients were also either positive or indeterminable by immunoblot testing. Histologic scores for parameters commonly associated with chronic viral hepatitis were numerically worse among anti-HCV-positive patients, but none reached statistical significance. Clinically, seven of 11 (64%) of anti-HCV-positive patients versus 14 of 26 (54%) anti-HCV-negative patients were Child's class C. Among the 21 Child's class C patients, seven (33%) were anti-HCV-positive versus four of 16 (25%) of Child's class A/B patients. A weak correlation between IgG and ELISA optical density was observed (r = 0.52). We conclude that antibody to hepatitis C by ELISA and immunoblot is common among alcoholics with liver disease even in the absence of known or suspected risk factors for viral hepatitis. Although hepatitis C-positive patients tended to have more severe histologic disease, neither histologic parameters nor clinical findings were adequate to predict antibody seropositivity.     

Molecular evidence that the hepatitis C virus replicates in the oral mucosa

BACKGROUND/AIMS: Patients infected with the hepatitis C virus (HCV) often have extrahepatic manifestations, which significantly contribute to HCV-related morbidity, but whose pathogenesis is largely unknown. Our aim was to evaluate the HCV replication in oral mucosa of chronic hepatitis C patients. METHODS: We collected oral mucosa specimens from 17 anti-HCV-positive and four anti-HCV-negative patients. Fifteen had oral lichen (12 anti-HCV-positive). Total mucosa RNA was extracted and analyzed for presence and titer of genomic and negative-strand HCV RNA. Findings were compared with clinical and pathological features. RESULTS: Genomic and negative-strand HCV RNA were detected, respectively, in 12 of 17 (70.6%) and four of 17 (23.5%) specimens from the chronic hepatitis C patients. No negative-strand HCV RNA was detected in five anti-HCV-positive patients without lichen (including three with normal mucosa). Presence and titer of the negative-strand HCV RNA were independent of HCV genotype, serum viral load, and histological diagnosis of liver lesions. The phylogenetic analysis of the envelope 2 region cloned from a normal mucosa and the corresponding serum further suggested that only lichen tissues appear to harbor replicating HCV. CONCLUSIONS: HCV may occasionally replicate in oral lichen tissue, possibly contributing to the pathogenesis of mucosa damage.     

Hepatitis-C-virus genotypes and hepatitis-G-virus infection in Lebanese thalassaemics

Exposure to hepatitis C virus (HCV), hepatitis G virus (HGV) and the carrier 'rate' for hepatitis B virus (HBsAg) were investigated in thalassaemia patients in Lebanon, a group that has not been studied in the past. The HCV genotypes and their distribution in the 395 thalassaemics, all of whom had been registered at the Chronic Care Center (CCC) in Hazmieh since 1996, were also studied. Of the 55 samples (14%) found positive for anti-HCV, 19 were also positive for HCV RNA. The 19 samples of HCV RNA were mostly of genotype 4 (37%), followed by 1a and 3a (21% each), lb (16%) and 2b (5%). Most (14; 74%) of the 19 HCV-RNA-positive samples, but only 13 (36%) of the 36 samples that were negative for HCV RNA although anti-HCV-positive, were positive for anti-HGV. Among 100 anti-HCV-negative samples, eight (8%) were anti-HGV positive. Only one (0.28%) of all 395 patients investigated was found to be HBsAg-positive. All of the HBV- and HCV-positive patients had initially been found positive in 1996, when they were first registered at the CCC, and none of the remaining patients had seroconverted since. As none of the patients had been checked for anti-HGV until the present study, the history of their exposure to HGV was unknown. These results emphasise the importance of screening all blood donations collected in Lebanon for HBsAg and anti-HCV. This and stringent infection-control measures are necessary steps to limit the spread of HBV, HCV and perhaps HGV to thalassaemics.     

Superinfection of TT virus and hepatitis C virus among chronic haemodialysis patients

BACKGROUND: The TT virus (TTV), a new DNA virus found in Japan from a patient with post-transfusion hepatitis non-A-non-G, is frequently positive in the sera of patients with liver disease. It is not established whether this virus causes liver damage. We studied the frequency of superinfection of this virus and hepatitis C virus (HCV) known to be endemic among haemodialysis patients, and the possible deleterious effect of TTV on HCV-induced chronic liver disease. METHODS: We used primers from a conservative region in the TTV genome (Okamoto, 1998) to detect TTV. Sera from 163 dialysis patients positive for anti-HCV and 77 dialysis patients negative for anti-HCV (control) were tested. RESULTS: TT Virus positivity was 35% among HCV antibody (anti-HCV)-positive patients and 45.4% among anti-HCV-negative patients. TT Virus positivity was unrelated to the length of haemodialysis or amounts of blood the patients had received in the past. More anti-HCV-positive patients had a history of transfusion, but TTV positivity was not as closely associated with transfusion as anti-HCV positivity. The severity of chronic liver disease was estimated from peak serum alanine aminotransferase levels in the preceding 6 months. Among anti-HCV positives, TTV-positive patients tended to have less active disease; at least there was no indication that TTV superinfection aggravated chronic hepatitic C in long-term dialysis patients. Four of 35 anti-HCV-negative, TTV-positive patients had chronic active liver disease, while none of the anti-HCV-negative and TTV-negative patients did. CONCLUSIONS: TT Virus infection is prevalent among haemodialysis patients. Its transmission occurs not only by blood transfusion, but also by non-parenteral infection. Superinfection of TTV does not exert deleterious effects on the liver disease induced by HCV. However, it may cause chronic hepatitis in a limited number of patients, but remains dormant most of the time. Triple infection, HCV and TTV plus HBV or HGV (one case each), did not cause severe liver disease.     

Hepatitis C virus infection in patients with leukemia

We have studied 30 patients with acute leukemia by the second-generation assay for antibodies to hepatitis C virus (HCV) to determine the incidence of HCV infection and the impact of anti-HCV positivity on liver disease. After a complete remission, 21/30 (70%) patients were anti-HCV-positive. During chemotherapy the anti-HCV-positive patients had more severe liver disease than the anti-HCV-negative patients, and they had a higher incidence of chronic hepatitis (13/21; 62% vs. 1/9; 11%, P < 0.01). During subsequent follow-up, 15/30 (50%) patients relapsed and 15/30 (50%) patients completed the chemotherapy protocols. After a relapse 12/15 (80%) patients were anti-HCV-positive and they had more severe liver disease than the anti-HCV-negative patients. Among the patients who completed chemotherapy (n = 15), biochemical evidence of chronic hepatitis was found in 9/9 (100%) anti-HCV-positive, and 2/6 (33%) anti-HCV-negative cases during off-therapy follow-up after therapy-withdrawal (P < 0.05). These results indicate that HCV plays an important role in the etiology of chronic hepatitis which could worsen the final prognosis of successfully treated patients with leukemia. © 1994 Wiley-Liss, Inc.     

Characterization of hepatitis C virus antibody positive blood donors in Japan and Thailand: a comparative study

The assay of specific antibodies to hepatitis C virus (anti-HCV) was introduced in the screening of donated blood in November 1989 in Japan and in January 1991 in Thailand. Anti-HCV-positive rates obtained using commercial second-generation kits on donated blood in both countries are similar. However we found differences in the serum alanine aminotransferase (ALT) values of anti-HCV-positive donors between the two countries: 57.0% of anti-HCV-positive Thai donors showed elevated ALT values, whereas only 23.7% of anti-HCV-positive Japanese donors did. Furthermore, different distributions of HCV genotypes were observed among anti-HCV-positive donors of the two countries. Although type 1b showed the highest prevalence among donors in both countries, type 1a showed the lowest prevalence in Japanese donors and the second-highest prevalence in Thai donors. We also examined the HCV RNA levels using a branched DNA probe assay in serum samples of Thai donors and observed no significant relationships between ALT value and HCV genotype or HCV RNA level, although HCV RNA levels in genotype 1b Japanese donors were higher than that in genotype 2a Japanese donors.     

Predictive Value of Screening Tests for Persistent Hepatitis C Virus Infection Evidenced by Viraemia: Japanese Experience

In November 1989, Japanese Red Cross Blood Centres started screening for heaptitis C virus (HCV) with enzyme-linked immunosorbent assay (Elisa) for the C100-3 viral peptide as the first such nationwide programme in the world. Thereafter post-transfusion non-A non-B hepatitis (PTNANBH) was reduced by 61–80%, but this was not as complete a success as our programme to prevent post-transfusion hepatitis B by screening for high titer hepatitis B core antibody, which we began in the same period. In order to acquire more effective control of PTNANBH, the HCV core-related antigen (GOR, N14) and second-generation Elisa (Ortho2, Abbott2)and second-generation antigen agglutination (PA, PHA) tests have been employed. Among 16,500 donors in 11 blood centers, 365 were serologically positive by at least one of these tests. Among these, HCV RNA was detected in 138 units and the remaining 227 were HCV RNA negatives. The effectiveness of these serological tests to detect HCV RNA-positive status were analyzed. Passive haemagglutination and particle agglutination (PHA and PA) tests were highly effective to predict HCV viraemia among blood donors. Also, these tests can easily determine antibody titre. By either PHA or PA, all units with ≧2¹² agglutination titre (120 and 122 units) were HCV RNA positive and all agglutination-positive units with serum alanine aminotransferase level higher than 35 Karmen units were HCV RNA positive. These results have formed the basis for implementing a more effective screening for HCV viraemia in blood donors, where effectiveness is defined as enhancing the protection of patients from post-transfusion hepatitis C and providing higher quality information to achieve more effective donor counselling.     

聚合酶链反应和杂交试验检测上海地区庚型肝炎病毒感染的初步报告

为探讨上海地区庚型肝炎病毒感染的现状，采用逆转录套式聚合酶链反应（ＲＴＮｅｓｔｅｄＰＣＲ）检测庚型肝炎病毒（ＨＧＶ／ＧＢＶＣ）核酸（ＨＧＶＲＮＡ）。结果在各类患者和助血员中ＨＧＶＲＮＡ的检出率分别是：血液透析和肾移植为１５７％（８／５１）、丙型肝炎为３３％（１／３０）、乙型肝炎为０（０／１９）、散发性非Ａ～Ｅ型肝炎为０（０／２８）、义务助血员为７５％（５／６７）。提示ＨＧＶ感染多为无症状或亚临床型，常与ＨＣＶ重叠感染，并与输血密切相关。作者肯定了上海地区存在庚型肝炎，指出筛选助血员对控制输血后庚型肝炎至关重要。     

Hepatitis C Virus (HCV) RNA among Anti-HCV-Positive Blood Donors and Their Recipients

Seven of 24 blood donors positive in Ortho's first-generation antibody to hepatitis C virus (anti-HCV) test (EIA-1) were also positive in Ortho's second-generation test (EIA-2). All 7 had at least two anti-HCV positive recipients, whereas only 1 of the 17 EIA-2-negative donors had an anti-HCV-positive recipient. This recipient was a multitransfused patient with von Willebrand's disease. Five of the 7 EIA-2-positive donors had detectable HCV RNA. We traced and tested 38 of the still living blood recipients from the 7 EIA-2-positive donors. Twenty-eight of these were EIA-2 positive and 22 were HCV-PCR positive. One patient with Waldenstroem's hypergammaglobulinemia was EIA-2 negative but HCV-PCR positive. All the EIA-2-positive sera showed reactivity in Ortho's recombinant immunoblot assay (RIBA-2), but 5 of the 28 recipients and 1 of the donors reacted with only one band (RIBA-2 indeterminate). Among 32 recipients who probably had received EIA-2-positive blood, 29 (91%) had markers of an HCV infection. Twenty-two (75%) of the HCV-infected recipients had detectable HCV RNA more than 6 months after transfusion and hence were chronic HCV carriers.     

Anti-HCV in post-transfusion hepatitis: deductions from a prospective study

Stored sera from 52 patients who developed post-transfusion hepatitis (PTH) during a prospective study of PTH in Toronto in 1984/85, sera from 111 donors whose blood was transfused into these patients and sera from 50 patients with chronic active hepatitis with a remote history of blood transfusion were tested for anti-HCV. In patients with PTH seroconversion occurred relatively early. Ten converted in less than 14 weeks after transfusion. Only three of the 34 patients (9%) whose hepatitis resolved developed anti-HCV compared to 11 of 18 (61%) whose hepatitis became chronic. Patients who seroconverted had higher alanine aminotransferase (ALT) values during the phase of acute hepatitis than those who did not seroconvert. Most of the patients who developed PTH received blood that was negative for anti-HCV. Four donors whose blood was positive for anti-HCV transmitted hepatitis. Three of the patients developed anti-HCV and chronic hepatitis. One of the recipients did not seroconvert and the hepatitis resolved. Forty-two of the 50 patients (84%) with chronic hepatitis and a remote history of blood transfusion were positive for anti-HCV. We conclude that anti-HCV-positive donors may transmit hepatitis C; that if anti-HCV is diagnostic of hepatitis C, most cases of acute PTH are either not due to hepatitis C or may represent cases of hepatitis C in which the anti-HCV test was undetectable. On the other hand, most cases of PTH which progress to chronic hepatitis are caused by HCV.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estimated Risk of Transmission of Hepatitis C Virus by Blood Transfusion

Objective : The risk of transmitting hepatitis C (HCV) by transfusion of anti-HCV-negative screened blood was estimated for the blood donor population of Baden-Württemberg (southwestern Germany). Methods : The data from the blood donors screened for anti-HCV and for HBsAg during 1990-1995 were analyzed. Results : The prevalence of confirmed anti-HCV-positive blood donations decreased continuously during the last 5 years, reaching 121 per 100,000 blood donations. A higher anti-HCV prevalence rate was found in female than in male blood donors (p<0.05). The estimated risk of transmitting HCV during the window period is 1:200,000 (1:97,000–1:1,400,000) for repeat donors. In 1995, the calculated risk for first-time donors was 1:20,000 (1:15,000–28,000). The incidence for HCV was 1.2 per 100,000 blood donations. Conclusion : The risk of transmitting hepatitis C by blood transfusion is low. Additional tests to shorten the window period to detect antibodies to HCV might increase the safety of blood transfusion.     

Hepatitis C

The major cause of chronic post-transfusion hepatitis, the hepatitis C virus (HCV), has been identified. HCV is a single-stranded linear RNA virus with characteristics similar to the flaviviruses. A different agent, the hepatitis E virus, is associated with epidemic (enterically-transmitted) non-A, non-B hepatitis. At present, infection with HCV is recognized by the finding of anti-HCV antibodies, positive in up to 90% of patients with chronic non-A, non-B post-transfusion hepatitis. Antibodies to HCV are detected in 1% of normal volunteer blood donors and in the majority of donors implicated in post-transfusion hepatitis. HCV antibodies are also found in patients with autoimmune liver disease and hepatocellular carcinoma. Moreover, HCV infection may contribute to the pathogenesis of liver disease in alcoholic patients. The role of HCV infection in fulminant non-A, non-B hepatitis and hepatitis-associated aplastic anemia has not been elucidated as yet. Therapy of chronic non-A, non-B hepatitis with recombinant human alpha-interferon has been shown to improve or normalize aminotransferase levels in approximately 50% of patients, most of whom have evidence of HCV infection. However, relapse after cessation of treatment is common. In the future, screening blood for evidence of HCV infection may prevent most cases of non-A, non-B post-transfusion hepatitis.     

A prospective study of transfusion-transmitted virus transmission by blood transfusion

Recently, a new single-stranded DNA virus (TT virus, TTV) has been isolated and related to post-transfusion hepatitis. The aim of this study was to investigate the prevalence of TTV in blood donors and blood recipients, and the incidence of TTV transmission by blood transfusion. TTV DNA and serum markers of hepatitis B virus (HBV) and hepatitis C virus (HCV), were examined in 130 blood recipients, and the presence of TTV was studied in their 340 corresponding blood donors. The prevalence of TTV infection was 10.6% (36/340) in donors and 8.5% (11/130) in blood recipients, before transfusion. Eighteen subjects (15.1%) were found to be TTV positive, after transfusion, in the 119 blood recipients without TTV before transfusion; at least one of the corresponding donors was TTV positive. There were 46 subjects with post-transfusion hepatitis virus infection, 45 with HCV infection (including seven co-infected with TTV) and two with HBV infection (including one co-infected with HCV and one co-infected with TTV). The recipient with TTV and HBV co-infection and three of the seven patients with TTV and HCV infection had alanine aminotransferase (ALT) levels higher than 90Ul^–1, but only two of the 10 isolated TTV infections had a mild ALT elevation. These results show that prevalence of TTV was high in blood donors and hospitalized patients, and isolated TTV infection is not related to significant ALT elevation.     

Also with a Restrictive Transfusion Policy,Screening with Second-Generation Anti-Hepatitis C Virus Enzyme-Linked Immunosorbent Assay Would Have Reduced Post-Transfusion Hepatitis C after Open-Heart Surgery

The incidence of post-transfusion hepatitis non-A, non-B (PTH-NANB) was prospectively assessed among open-heart surgery patients from the southeast region of Sweden before the introduction of anti-hepatitis C virus (HCV) blood donor screening. Blood samples for alanine aminotransferase analysis were drawn before and 2, 3, and 4 months after transfusion. Surgery was performed in four centres. Of 190 transfused and followed-up patients 2 (1.1%) contracted PTH-NANB, both operated on at the centre with significantly fewer transfusions than the other centres. One patient had antibodies to HCV detected by first-generation (C100-3) and later by second-generation anti-HCV enzyme-linked immunosorbent assay (ELISA-2) and by positive second-generation recombinant immunoblot assay (4-RIBA). The other patient, although negative by first-generation anti-HCV ELISA, was positive by second-generation ELISA and by 4-RIBA. Both patients were hepatitis C-viremic by polymerase chain reaction (PCR). All the six donors implicated in the two hepatitis cases were first-generation anti-HCV-negative, but two, one for each patient, were positive by second-generation anti-HCV ELISA. This finding was confirmed by positive 4-RIBA in only 1 donor, the other being ‘indeterminate’. However, in both donors hepatitis C viremia was found by PCR. This study shows that the second-generation anti-HCV ELISA will further reduce the risk for PTH-NANB/C and draws attention to the problem of evaluation of confirmatory tests.     

HCV infection and diabetes mellitus: influence of the use of finger stick devices on nosocomial transmission

An increased prevalence of hepatitis C virus (HCV) infection in patients with diabetes mellitus has suggested a link between these two conditions and the possibility of patient-to-patient HCV transmission during hospital admissions in diabetes units. We investigated the prevalence of HCV antibodies in 259 patients with diabetes mellitus consecutively admitted to our diabetic unit in 1998. The control group was composed of 14,100 volunteer blood donors. We divided the diabetic patients into two groups according to their HCV antibody status and also analysed patients for the following variables: age, disease duration, diabetes treatment, previous hospital admissions in a diabetes unit and use of finger stick devices. Anti-HCV antibodies were detected in 8 diabetic patients and 6 blood donors (3.09% vs 0.04%, p < 0.001). No differences were observed between anti-HCV-positive and anti-HCV-negative diabetic patients in terms of mode of treatment, previous hospital admissions in a diabetic unit and use of finger stick devices for capillary blood sampling. Our findings indicate that these medical practices play no role in nosocomial transmission of HCV in diabetic patients.