Prevalence of false-positive hepatitis C antibody results,National Health and Nutrition Examination Study (NHANES) 2007–2012

BackgroundScreening large numbers of persons in a population with low prevalence of a disease leads to many false-positives. However, populations with low HCV prevalence may sometimes be recommended for HCV screening, for instance patients or healthcare workers after a possible healthcare-related exposure.ObjectivesWe determined the percentage of true vs false-positive HCV antibody (anti-HCV) test results among 2007–2012 participants in the National Health and Nutrition Examination Study (NHANES), a nationally representative study with approximately 1% HCV infection prevalence, much lower than in groups typically recommended for HCV screening.Study designAnti-HCV test confirmation was performed using a recombinant immunoblot assay (RIBA) test and follow-up HCV RNA testing.ResultsOverall, of 22,359 NHANES participants tested, 479 (2%) were anti-HCV screening reactive and 477 were tested for RIBA; of these 323 (68%) confirmed as true positive and 105 (22%) were false-positives. Many others (49, 10%) were RIBA indeterminate and likely false-positive. Because of these false positive tests, the overall prevalence of chronic infection among those testing anti-HCV screening reactive was much lower (218, 51%) than would be expected due to disease clearance alone (approximately 80%).ConclusionsAll screening anti-HCV positive tests should be followed by an HCV RNA test, in order to confirm whether the patient has current infection so that infected persons can be referred to care and treatment to avoid the significant morbidity and mortality associated with chronic HCV infection.     

High prevalence and incidence of hepatitis C virus infections among dialysis patients in the East-Centre of Tunisia

Hemodialysed patients are recognised as a group at increased risk of infection with hepatitis C virus (HCV). The aim of this study was to determine the prevalence and incidence of HCV infection among dialysis patients of the east-centre part of Tunisia. Two hundred and seventy-six patients dialysed until 2001 were recruited within seven hemodialysis units located in the cities of Sousse, Monastir and Mahdia. The serum markers of HCV infection were tested over the period of March 2000-December 2002, by a 3rd generation ELISA test for antibodies and by qualitative RT-PCR technique for viral RNA. The prevalence of anti-HCV antibodies and of HCV RNA was 32.6% (90 patients) and 25.7% (71 patients), respectively. Between 1998 and 2002, 20 new infections were documented in five of the seven dialysis units corresponding to an incidence of 2.34% per year, with an average time of contamination after the beginning of dialysis of 4.6 years. If all the infections are assessed to have occurred during dialysis, the density of incidence of HCV contamination was 4.4% per year of dialysis. A high correlation was noticed between the presence of HCV markers in serum and the duration of dialysis (F = 34.15, P < 0.0001). In the absence of other risk factors (transfusion, drug-addiction), these results plead for the nosocomial transmission of the observed HCV infections. A phylogenetic analysis of the E2 hypervariable region of the viral genome is in progress to confirm this assumption.     

Incidence of HCV infection in French hemodialysis units: a prospective study

A large prospective study was carried out from 1997 to 2000 in 25 French hemodialysis units including 1,323 patients to determine the incidence of hepatitis C virus (HCV) infection. Monthly testing of alanine aminotransferase (ALT) activity, and assessment of HCV RNA and anti-HCV antibodies if the ALT activity was elevated, identified 14 new infections in 7 different units, giving an incidence of 0.4% new HCV infections per year. Molecular analyses and epidemiological data indicated that five patients became infected with HCV outside the unit where they were dialyzed, while the nine remaining patients acquired HCV from infected patients on dialysis during the same shift at the same unit. HCV was cleared in six of the seven (85.7%) patients with acute hepatitis C who were given standard doses of alpha-interferon (alpha-IFN). The persistence of nosocomial transmission of HCV in hemodialysis units emphasizes the need to implement infection control practices. Identifying new infections is crucial because alpha-IFN treatment results in long term clearance of HCV RNA in a large proportion of patients.     

Molecular epidemiology of a hepatitis C virus outbreak in a hemodialysis unit in Italy

Hemodialysis patients are at increased risk of hepatitis C virus (HCV) infection. The aim of this study was to investigate a HCV outbreak in a hemodialysis unit using epidemiological and molecular methods. Between April 2003 and October 2003, anti-HCV seronconversion was detected in four patients attending the unit. These cases were added to 10 patients already anti-HCV positive upon admission in the unit. All 14 anti-HCV patients were tested for HCV RNA and HCV genotype. NS5B and HVR1/ E2 genomic regions were amplified and sequenced in all HCV RNA positive patients and phylogenetic analysis was performed. Furthermore, clinical-epidemiological records obtained from all patients were examined. All four patients newly infected harbored genotype 2c. Genotype 2c was also detected in 2 of 10 patients already anti-HCV positive upon admission. Phylogenetic analysis showed that all newly HCV infected patients harbored very closely related viral isolates that clustered together with the 2c isolate found in one of the two 2c chronic infected patients. All HCV-2c infected patients had no other risk factors except hemodialysis. Three of four newly HCV-2c infected patients and the one HCV-2c chronically infected involved in the outbreak received dialysis on the same day and same shift but used different machines. The remaining HCV-2c newly infected patient and one of the above cited three received dialysis on the same day during different shifts but used the same machine. The outbreak was probably due to breaks of infection control procedures although a related-machine transmission cannot be excluded in one of the cases.     

Detection of hepatitis C virus sequences in sera with controversial serology by nested polymerase chain reaction.

The specificity of first-generation enzyme-linked immunosorbent assays (ELIAs) for antibody detection in individuals with hepatitis C virus (HCV) infection has been questioned in some pathological situations. We observed a surprisingly high prevalence of anti-HCV antibodies in alcoholic patients, and thus, false-positive reactions in anti-HCV tests were strongly suspected. The introduction of new epitopes, particularly a core protein, C22 (second-generation tests), seems to increase the sensitivity of anti-HCV detection. In order to study the specificity of the second-generation tests, 60 serum samples from alcoholic patients found to be positive by the first-generation anti-HCV ELISA (Ortho) were reexamined by a second-generation anti-HCV enzyme immunoassay (Abbott) and a recombinant immunoblot assay (RIBA II; Chiron). Fifteen serum samples gave contradictory results when they were tested by the two assays. We performed nested polymerase chain reactions (PCRs) to confirm that the discrepancies that we observed could be due to the presence of low levels of anti-HCV antibodies, which were detected by a more sensitive test, or to unspecific positive reactions. Nested PCR revealed the presence of HCV RNA sequences in all anti-HCV-positive sera or sera that were weakly positive by ELISA. Anti-HCV positive by RIBA II was always correlated with the presence of viral RNA in serum, but HCV RNA was detected in RIBA II-negative sera. These results indicate that the specificity of the second-generation tests is an important improvement but that an HCV infection can still persist without detectable antibodies. PCR remains the reference assay to clear up controversial serology results and to detect HCV infection in patients with no anti-HCV-detectable immune response.     

Low prevalence of anti-hepatitis C virus antibodies in female hemodialysis patients without blood transfusion: A multicenter analysis

To investigate whether nosocomial infection with hepatitis C virus (HCV) in chronic hemodialysis patients is related primarily to hemodialysis procedures, a multicenter analysis was carried out on 2,132 chronic hemodialysis patients (male: 1,274, female: 858) from 23 dialysis units using a second-generation anti-HCV antibody assay. The prevalence of anti-HCV antibodies in patients with blood transfusion (29.9%) was significantly higher (P < .0001) than in those without blood transfusion (7.6%). Although the prevalence of anti-HCV antibodies increased with the length of hemodialysis in males without blood transfusion, it did not increase even after long-term hemodialysis (more than 5 years) in females without blood transfusion, who exhibited a rate (1.9%) similar to that of healthy blood donors in Japan. There was a significant correlation between the presence of anti-HCV antibodies and anti-HBs antibody in males without blood transfusion. In anti-HBs antibody-negative male patients without blood transfusion, the prevalence of anti-HCV antibodies was significantly lower compared with anti-HBs antibody-positive male patients without blood transfusion. There was marked difference in the prevalence rate in patients without blood transfusion among dialysis units, and there was no correlation between the prevalence and the mean period of dialysis of each dialysis unit. Although nosocomial infection with HCV appears to be related to the hemodialysis environment, the low prevalence of anti-HCV antibodies in females suggests that dialysis procedures per se may not present the risk of hepatitis C virus infection. © 1996 Wiley-Liss, Inc.     

Prevalence of hepatitis C virus antibodies among patients infected with human immunodeficiency virus.

A study was undertaken to determine the prevalence and risk factors for serological evidence of hepatitis C virus (HCV) infection in patients infected with the human immunodeficiency virus (HIV). Tests for anti-HCV antibody were carried out by enzyme-linked immunoassay (EIA) on 101 HIV-infected patients from two university-based outpatient clinics. Anti-HCV antibody reactive samples were tested by using a recombinant immunoblot assay (RIBA) for HCV antibodies. Fourteen of 101 (13.9%) HIV-infected patients were anti-HCV reactive by EIA. Of these 14, only seven were reactive by RIBA: four were intravenous drug users as a sole risk factor for HIV infection; and the remaining three acquired HIV by blood transfusion, contaminated instrument exposure or IV drug use and sexual contact. Acquisition of HIV by sexual activity alone was not associated with HCV infection. It is concluded that HCV infection is found in approximately 7% of a university HIV clinic population. False-positive anti-HCV antibody serology may lead to overestimation of the prevalence of HCV infection. Female sex and intravenous drug use are significantly associated with HCV infection among HIV-infected individuals.     

Determination of hepatitis C virus genotypes among blood donors in Ahvaz,Iran

This study aims to determine the genotypes of hepatitis C virus (HCV) among blood donors at Ahvaz Blood Transfusion Centre. Blood samples were taken from 2376 blood donors -$$$ 1795 (75.54%) male and 581(24.45%) female -$$$ who referred to Ahvaz Blood Transfusion Centre during 2007-2008. Detection of anti-HCV antibody for all the donors was carried out by ELISA and the confirmatory RIBA tests. HCV RT-PCR followed by RFLP test was carried out for anti-HCV positive samples. Out of 2376 blood donors, only 55 (2.3%) male donors showed to be positive for HCV antibody by ELISA and RIBA tests out of which 45(1.8%) donors were positive for RT-PCR test. Female donors were negative for HCV antibody. The result of HCV genotyping by RFLP test showed 24 (53.3%) for 1a, 17 (37.7%) for 3a (α) and 4 (8.8%) for 3a (β) genotypes respectively. In conclusion, high prevalence of 53.3% HCV 1a genotype was observed among blood donors in Ahvaz city.     

Hepatitis C virus infection in Jordanian haemodialysis units: serological diagnosis and genotyping

The seroprevalence and genotypes of hepatitis C virus (HCV) were studied in 283 patients attending six haemodialysis units in Jordan. In all, 98 (34.6%) patients were anti-HCV-positive by EIA, 92 (93.9%) of whom were also reactive in an immunoblot assay. The prevalence of anti-HCV was correlated with a history of blood transfusion before the introduction of blood donor screening for HCV and with duration of haemodialysis. HCV RNA was detected in 30 (30.6%) of 98 anti-HCV-positive sera. HCV viraemia was not associated with a particular antibody for the six HCV antigens studied by the immunoblot assay, although reactivity to the core antigens was greater in the HCV RNA-positive sera than in negative sera. Two HCV genotypes (1 and 4) were identified for the first time in Jordan by restriction fragment length polymorphism analysis of HCV 5'-NCR. The predominant genotype was HCV la (12 of 30). Genotypes lb and 4 were detected in 10 and 8 patients, respectively. The antibody response to HCV antigens was genotype-dependent, with a wider range of antibody specificities detected in the immunoblot assay in the 12 patients with genotype 1a infection than in the 8 patients with genotype 4. However, there was no significant difference in the prevalence of antibodies to HCV antigens among patients infected with either genotype 1a or lb. In conclusion, the prevalence of anti-HCV, blood transfusion, duration of dialysis and HCV genotypes suggest possible nosocomial HCV transmission among patients which needs confirmation by phylogenetic analysis of subgenomic HCV regions.     

Low prevalence of antibody to hepatitis C virus in north east England

The prevalence of antibodies to hepatitis C virus (anti-HCV) was studied in North East England in blood donors, local multiply transfused patients, local high risk individuals, and chronic liver disease patients. Anti-HCV was detected by enzyme-linked immunosorbent assay (ELISA) in 2/1120 (0.18%) blood donors; 1/84 chronic renal failure patients on haemodialysis who had received 1,992 units of blood (seroconversion rate of 0.05% per unit transfused), 1/207 cardiac patients 6 months post cardiac surgery transfused with 1,403 units of blood (1 anti-HCV pre-operatively, seroconversion rate 0.07%), 40/50 haemophilia A patients treated with commercial factor VIII, and 38/100 intravenous drug users. In addition anti-HCV was detected by ELISA in 5/35 cryptogenic chronic liver disease patients, 5/5 confirmed by recombinant immunoblot assay (RIBA) (14%); 3/30 patients with autoimmune chronic active hepatitis, 2/3 by RIBA (7%); 2/50 primary biliary cirrhosis patients, 1/2 by RIBA (2%); 0/30 alcoholic cirrhosis patients; and 2/9 patients with hepatocellular carcinoma, 1/2 by RIBA (11%). HCV is uncommon in North East England; it may be implicated in the aetiology of a minority of cases of cryptogenic liver disease and less than 5% of autoimmune chronic active hepatitis and primary biliary cirrhosis.     

Monitoring hepatitis C infection in a major Swedish nephrology unit and molecular resolution of a new case of nosocomial transmission

Hepatitis C virus (HCV) infection is a frequent problem in hemodialysis units. The prevalence and incidence of HCV infection over a decade were studied in a nephrology unit affected by previous nosocomial HCV transmission. The HCV non‐structural 5B protein gene was sequenced to achieve phylogenetic analysis of a new (incident) case of infection. Proportions of patients who were and were not infected with HCV remained similar over the period, as did the inflow and outflow of patients infected previously. In 1997, 12/157 (8%) of patients at the unit (treatment: hemodialysis, peritoneal dialysis, and renal transplant recipients) were positive in HCV RNA, whereas in 2007 the overall number was 9/239 (4%). One patient acquired an HCV infection, and the NS5B sequence in that case clustered with genotype 2b sequences found in patients from an earlier outbreak. Comparing the HCV from the incident patient with several stored longitudinal samples and cloned PCR products from the most likely source patient revealed close phylogenetic relationship with an HCV quasispecies member from the possible source. The source patient and the incident newly infected patient were not scheduled on the same dialysis shift, although the records showed that simultaneous treatment occurred on two occasions during the months preceding transmission. In conclusion, over the 10‐year period, the proportion of HCV‐infected patients at the unit was unchanged. Only one new infection occurred, which originated from a fellow patient's quasispecies. This establishes phylogenetic analysis as a valuable tool for tracing patient sources of HCV transmission. J. Med. Virol. 82:249–256, 2010. © 2009 Wiley‐Liss, Inc.     

Prevention of nosocomial transmission of hepatitis C infection in a hemodialysis unit. A prospective study

Hepatitis C virus (HCV) infection in hemodialysis patients can be transmitted by transfusions and nosocomially. A high prevalence of HCV infection, over 50%, was demonstrated in our hemodialysis (HD) unit. In order to prevent the nosocomial spread of HCV infection in the HD unit a prospective study was begun separating anti-HCV positive patients from the negative ones. A total of 170 patients (83 anti-HCV positive) started this study in September 1994 and were followed for 4 years. A separate room and dedicated equipment were assigned to anti-HCV positive and anti-HCV negative patients. Of those 170 patients there were 15 hepatitis B virus (HBV) positive patients, 5 of whom were anti-HCV positive, who were treated in a separate room on dialysis equipment for anti-HCV positive or negative patients. Application of general precautions, as recommended by the Center for Disease Control and Prevention (CDC), was reinforced. During the first 12 weeks after implementing the precautions seven more anti-HCV positive patients were detected, and by December 1995 another two HCV infected patients were found. The follow-up included all changes in HD population treated until the end of 1998. The incidence of seroconversion to HCV was 12.9% in 1995, 7.1% in 1996, 5.0% in 1997, and 6.6% in 1998. The higher incidence of seroconversion in September to November 1994 was probably due to the nosocomial infection being in the incubation period at the time of isolation. This prospective study in a large HD unit with a high prevalence of HCV infection demonstrates a relatively successful prevention of HCV spread. Procedure-related transmission of HCV in hemodialysis could be prevented by rigorous application of universal precautions as recommended by the CDC. As a second line of prevention, in highly burdened dialysis centers, segregation of HCV positive patients can help control nosocomial transmission.     

Hepatitis C virus infection among pregnant women in Yaounde,Cameroon: prevalence,viremia, and genotypes

Central Africa is considered to be an area of high endemic hepatitis C infection. To determine the prevalence of anti-HCV antibodies, HCV RNA, and the genotype distribution in Cameroon, 1,494 pregnant women attending antenatal care units in Yaounde, Cameroon were screened for HCV infection. Anti-HCV antibodies were detected with a 3rd generation ELISA (Monolisa anti-HCV plus version 2, BioRad, Richmond, CA). All anti-HCV antibody-positive sera were then tested with another 3rd generation ELISA (AxSYM) HCV version 3, Abbott Laboratories, Abbott Park, IL) and subsequently for HCV RNA (Amplicor HCV, Roche Diagnostics, Basel, Switzerland). Genotype was determined by phylogenetic analysis of the NS5b gene. Seventy-three pregnant women were found to be anti-HCV antibody positive by the first ELISA, but only 28 were anti-HCV positive by both ELISA. The prevalence of anti-HCV antibodies was thus 1.9% (28/1,494) (95% CI: 1.3-2.7%). 21/28 (75%) of the positive samples by both ELISA were HCV RNA positive. The 45 samples that were HCV antibody negative by the second ELISA were also HCV RNA negative. The HCV subtypes identified were 1a (24%), 2f (38%) and 4f (38%). In contrast to previous studies, anti-HCV antibodies were rare among pregnant women in Cameroon. The percentage of HCV seropositive pregnant women who had circulating HCV RNA was similar to that observed in Europe. Several HCV genotypes were found in Cameroon.     

Prevalence of anti-HCV and HCV viremia in hemodialysis patients in Taiwan.

Hepatitis C viral infection in 125 hemodialysis patients from Taiwan was studied using a second-generation anti-HCV immunoassay (EIA II) (Abbott HCV 2.0 EIA) and the polymerase chain reaction (PCR) to detect the HCV RNA in the serum. A total of 59 patients (47.2%) were positive by EIA II. In comparison, the conventional C100-3 anti-HCV assay was positive in 40 (32.0%). HCV RNA was found in 47 patients (37.6%). Patients with elevated serum transaminase level had a higher positive rate of anti-HCV and HCV RNA. The dialysis time was longer for those patients positive for anti-HCV than for those who were negative. A total of 57 of the 59 EIA II-positive cases had a history of blood transfusion. The HBsAg status did not influence the anti-HCV positivity. Among the 59 EIA II-positive patients, 66.1% were also positive for HCV RNA, and of the 47 HCV RNA-positive cases 83.0% were positive for EIA II. It is concluded that the high prevalence of specific HCV infection and HCV viremia was present in these patients. Prevention of cross-contamination during dialysis and blood screening before transfusion are important for the control of HCV infection in these patients.     

Use of phylogenetic analysis of hepatitis C virus (HCV) hypervariable region 1 sequences to trace an outbreak of HCV in an autodialysis unit

Hemodialysis patients are at high risk of infection by hepatitis C virus (HCV). The aim of this study was to investigate an HCV outbreak that occurred in an autodialysis unit by using epidemiological and molecular methods. Seroconversion to HCV antibody (anti-HCV) was observed in two patients over an 18-month period; two other patients had previously been recorded as anti-HCV positive. All four patients involved in the outbreak were tested for HCV RNA, and hepatitis C genotype determination was accomplished by a reverse hybridization assay. Furthermore, part of hypervariable region 1 (HVR1) of the hepatitis C genome was amplified and sequenced in samples from all HCV RNA-positive patients. Phylogenetic analysis of the nucleotide sequences obtained was carried out in order to investigate any possible epidemiological linkages among patients. The nucleotide sequences of the HVR1 regions of both newly infected patients were found to be identical to sequences of samples from previously recorded anti-HCV-positive original patients, suggesting that they were infected by the same isolate. Molecular and epidemiological analysis suggested that nosocomial patient-to-patient transmission was the most likely explanation for the virus spread in the autodialysis unit under study.     

Significance of indeterminate third-generation hepatitis C virus recombinant immunoblot assay.

Indeterminate hepatitis C virus (HCV) third-generation recombinant immunoblot assay (RIBA3.0; Ortho Diagnostic Systems) patterns were arbitrarily defined by the manufacturer as the detection of only one antibody out of the four that were sought, namely, c100 (NS4 encoded), c22 (core encoded), c33c (NS3 encoded), and NS5 (NS5 encoded). The aims of the present study were (i) to determine the prevalence of indeterminate RIBA3.0 patterns in patients consecutively tested for anti-HCV antibodies in a university hospital; (ii) to evaluate the significance of these patterns in terms of viral replication, liver disease, and risk factors for HCV; and (iii) to get an insight into the mechanism underlying this peculiar immune response. Among 3,074 serum samples consecutively tested for anti-HCV antibodies, 588 were found to be positive by screening assays. Fifty-nine of them (10%) were RIBA3.0 indeterminate and were compared with 59 RIBA3.0-positive ones. Thirty-one RIBA3.0-indeterminate and 53 RIBA3.0-positive serum samples were HCV RNA positive by PCR (53 versus 90%; P < 10(-6). RIBA3.0-indeterminate and RIBA-3.0-positive patients with positive PCR results were not significantly different for the prevalence of risk factors for HCV infection and elevated serum alanine aminotransferase activities. Immunosuppression, attributable to coexisting human immunodeficiency virus infection, organ transplantation, or the administration of immunosuppressive drugs, was significantly more frequent in PCR-positive, RIBA3.0-indeterminate patients than in PCR-negative, RIBA3.0 indeterminate patients (P < 0.001) and PCR-positive patients with a positive RIBA3.0 result (P < 0.01). The distribution of HCV genotypes did not differ significantly between HCV RNA-positive patients with indeterminate or positive RIBA3.0 results. In conclusion, the prevalence of indeterminate RIBA3.0 patterns in virology laboratories is about 10%; in about half of these patients HCV replication is detected by PCR; the main factor responsible for indeterminate RIBA3.0 patterns could be immunosuppression, whereas HCV genotypes do not seem to play major role.     

Prevalence of hepatitis C virus infection among hemodialysis patients at a tertiary-care hospital in Mexico City, Mexico

We determined the prevalence of hepatitis C virus (HCV) in hemodialysis patients by antibody testing and HCV RNA determination by PCR. A total of 149 patients with kidney failure with replacement therapy were tested. The prevalence of anti-HCV was 6.7% (10 of 149 patients), and viremia was detectable in 8 of 149 (5%) patients. Three of 149 patients (2%) were anti-HCV negative with detectable HCV RNA.     

Evolution of hepatitis C virus (HCV) infection acquired at birth

Objectives Our aim was to analyze the evolution of HCV infection in children infected at birth. Methods Between September 1994 and December 1998 we analyzed in a prospective study 8 children born of anti-HCV and HCV RNA positive women. Each baby was controlled at birth, every 3 months during the first year of life, and then every 6 months searching for anti-HCV antibodies (ELISA 3, RIBA 2-3), HCV RNA (RT PCR), ALT and viral genotype. Results Viral RNA was detectable in the first 3 months of life in all babies (100%) and remained positive during the follow-up. Viral genotypes were the same for mothers and their children. In 6 babies (75%) ALT remained pathologic during follow-up. Conclusions HCV infection in children usually has an asymptomatic outcome; the infection has chronic features in the majority of cases.