anti-HCV Negative Patients Research Articles

The prevalence of HCV RNA positivity in anti-HCV antibodies-negative hemodialysis patients in Thrace Region. Multicentral study.

HCV infection in patients under hemodialysis for end stage chronic kidney disease (ESCKD) may exist despite the absence of anti-HCV antibodies. Molecular methods are widely accepted as "gold standard" techniques for the detection of viral RNA. However, the molecular methods are more expensive in comparison to conventional methods and their replacement is not cost-effective. The aim of this study was to estimate the prevalence of HCV RNA positivity in anti-HCV negative hemodialysis patients and evaluate new diagnostic methods for the detection and the monitoring of hepatitis C in ESCKD patients. The study was performed in four hospitals of Thrace region of Greece and 233 patients with no history of hepatitis C were enrolled. Measurement of anti-HCV antibodies and HCV core antigen was performed by microparticle chemiluminescence immunoassay. Molecular detection of viral RNA was performed by the real-time RT PCR. The mean age of the patients was 64.9 ± 23.3 years. HCV-Ag was positive in 2/233 patients (0.86%). Nevertheless, viral RNA was negative in those patients. The results of the present study showed that the incidence of HCV-RNA in patients with negative anti-HCV Abs, in hemodialysis patients in Thrace region of Greece was negligible (0/233).

High Seroprevalence of Hepatitis C Virus Antibody in Breast Cancer Patients in Egypt.

Background:Hepatitis C virus (HCV) is a known risk factor for hepatocellular carcinoma. Several epidemiological studies have pointed out to an association of HCV infection with other extrahepatic malignancies. The role of chronic HCV in breast cancer causation is less clear. Egypt is an endemic area of HCV infection with resulting significant morbidity. The association between HCV status and breast cancer risk in Egyptian women is hitherto unknown.Methods:A retrospective study was performed. The prevalence of anti-HCV seropositivity was estimated in a sample of women with a breast cancer diagnosis, retrieved from the hospital records, and was compared to the raw data of a population study in Egypt. Anti-HCV negative and positive patients were compared regarding the disease course and outcome.Results:Retrospective analysis revealed a markedly high prevalence of anti-HCV seropositivity in young breast cancer patients. In patients younger than 45 years, 13.4% were anti-HCV positive. Seropositivity was 6-fold higher in these patients than in adult females of the same age without cancer diagnosis (P = .003). The biological type, tumor size, nodal status, and disease-free survival were not affected by the patients’ HCV status.Conclusion:Young Egyptian breast cancer patients have a dramatically high prevalence of HCV seropositivity. Further population studies are strongly required to investigate the epidemiological association of these two significant health problems.

Genotyping and prevalence of hepatitis c virus in dialysis patients in Gandhi general hospital - A tertiary care centre

Context: The present study was undertaken to determine the genotyping and prevalence of hepatitis c virus in dialysis patients as HCV genotyping has become a part of the pre-treatment evaluation. Studies correlating risk factors and genotypes of HCV infection prevalent in Haemodialysis patients are scanty. Therefore our study is conducted to study the prevalence, genotypes and risk factors in order to detect and try to curb HCV infection. Aims: To study the genotypes of hepatitis C infection and its prevalence in hemodialysis patients. Settings and Design and Period: Study was conducted in Gandhi hospital Secunderabad in 1 year period as Cross-sectional study. Materials and Methods: A total of 225 serum samples were collected from chronic renal failure patients who were undergoing Haemodialysis over a period of 1 year. The samples were screened for anti HCV antibodies by third generation ELISA test. 25 anti-HCV positive samples and 40 anti-HCV negative samples were randomly selected for HCV RNA detection using Trizol-Chloroform-Isopropyl alcohol method, HCV RNA was reverse transcribed with 200 U of maloney murine leukemia virus reverse transcriptase and random hexamers. Nested PCR was done. HCV RNA positive samples were genotyped with primers specific for core region of different HCV types by type specific PCR. Statistical Analysis Used: Statistical analysis of the data was done by chi-square (c2) test using EPIINFO 2000 software. The values of chi square test are interpreted as not significant >0.05, highly significant < 0.001. Results: Out of 225 hemodialysis patients 38 (16.8%) patients were anti HCV positive. Duration of dialysis was significantly longer in anti-HCV antibodies positive group with dialysis duration more than 2 years. Seropositivity is more in HD patients having dialysis at more than one centre. HCV RNA was detected in randomly selected 13/25 (52%) anti HCV positive patients and in 5/40 (12.5%) anti HCV negative patients. The genotype distribution

Seroprevalence of Hepatitis B and Hepatitis C in end-stage renal disease patients on maintenance hemodialysis: a single-center study from Bangladesh

Background: Hepatitis B virus (HBV) and Hepatitis C virus (HCV) infections are important causes of morbidity and mortality in hemodialysis (HD) patients. The aim of this study was to estimate the seroprevalence of HBV and HCV among end-stage renal disease (ESRD) patients on maintenance HD.Methods: This cross-sectional study was conducted in the hemodialysis unit of a military hospital of Bangladesh from October 2013 to March 2014 and included 141 maintenance HD patients. All patients were assessed by HBsAg and anti-HCV antibodies in addition to routine liver function tests.Results: The age range of the study subjects were 18-70 years, and the majority (85.11%) were male. Among them 22 (15.60%) were positive for anti HCV, 5(3.5%) were positive for HBsAg, and 3 patients (2.13%) were positive for both HBsAg and anti-HCV. The duration of hemodialysis was higher in anti-HCV positive patients (49±24 vs. 25±10 months, p &lt;0.05) than anti-HCV negative ones. Anti-HCV positive patients in this study received a higher number of blood transfusion (units) than anti-HCV negative patients (7.5±4.3 vs. 2.8±1.7 units, p &lt;0.05). HBsAg positive patients also received a higher number of blood transfusion (units) than HBsAg negative patients (8.2±3.1 vs. 3.2±1.2 units, p &lt;0.05).Conclusions: Hepatitis C virus was the major form of hepatitis in HD patients in this study. The duration of HD was higher in anti-HCV positive patients; the numbers of blood transfusion units were higher in patients positive for HCV and HBV than the negative ones.

Existence of Occult Hepatitis C virus Infection in Local Haemodialysis Setting

Introduction: Among chronic hepatitis C patients, the viral agent is well known to replicate in the hepatocytes and extrahepatic cells. Previous studies have demonstrated a new form of hepatitis C called 'Occult HCV Infection' (OCI). It is defined as the presence of HCV RNA in the hepatocytes but undetectable HCV RNA in the serum or plasma. The exact natural history, epidemiology, pathogenesis and clinical importance of OCI are unknown. Several studies have been carried out worldwide, but none from Malaysia. This study investigated OCI among local patients undergoing routine hemodialysis at a referral hospital in Pahang, Malaysia. Materials and Methods: Blood was collected from both anti-HCV positive (n=10) and anti-HCV negative (n=18) patients. Inclusion criteria for the seronegative patients included elevated liver enzymes. Results: 10 chronic hepatitis C patients were recruited in this study. All had viral RNA detected in both serum and peripheral blood mononuclear cells (PBMC). Viral quasispecies was common in this group of patients. Meanwhile, out of 18 seronegative patients, 3 patients (16%) showed active viral replication in PBMC. Conclusion: This study proved the existence of OCI in local haemodialysis setting. Further investigation is needed to get the actual prevalence of OCI.

The efficacy of direct anti-HCV drugs improves early post-liver transplant survival and induces significant changes in waiting list composition

The efficacy of direct-acting antivirals (DAAs) has dramatically changed the prognosis of patients with chronic hepatitis C. We aimed to evaluate the impact of DAA therapy on the composition of the liver transplant (LT) waiting list and the early post-transplant survival. We evaluated all patients admitted to the waiting list for a primary LT between 1st January 2008 and 31st of December 2016 in Catalonia, Spain. Time span was divided into two periods according to the availability of different antiviral therapies: 2008-2013 (interferon-based therapies) and 2014-2016 (DAA). Changes in the indications of LT and the aetiology of liver disease, as well as post-LT patient survival, were evaluated according to the year of inclusion and transplantation, respectively. We included 1,483 patients. Admissions in the waiting list for hepatitis C virus (HCV)-related liver disease decreased significantly, from 47% in 2008-2013 to 35% in 2014-2016 (p <0.001), particularly because of a reduction in patients with decompensated cirrhosis. In contrast, NASH-related inclusions increased from 4% to 7% (p = 0.003). Three-year post-LT patient survival increased significantly in the second period in the whole cohort (82% vs. 91%, p = 0.002), because of better survival in anti-HCV positive patients (76% vs. 91%, p = 0.001), but not in anti-HCV negative patients (88% vs. 91% p = 0.359). Anti-HCV positive serology, the time period of 2008-2013 and higher donor age were independently associated with post-LT mortality in the whole cohort; while time period and donor age were independently associated with post-LT mortality in anti-HCV positive recipients. The high efficacy of DAAs is associated with significant changes in the composition of the LT waiting list and, more importantly, results in improved post-transplant survival. The efficacy of the new direct-acting antivirals is associated with a significant improvement in survival of patients undergoing liver transplantation because of hepatitis C virus-related liver disease. In addition, it has decreased the number of patients with hepatitis C that need a liver transplant.

Seroprevalance of hepatitis-c infection in multi-transfused thalassemic children: study from a West Indian tertiary care center

Background: To study the prevalence of hepatitis-C virus infection in multi-transfused thalassemic children and to correlate these patients with age, number of transfusion, serum ferritin levels and transaminases levels.Methods: This study was conducted in the Department of Pediatrics of a Teaching Institute of Rajasthan. It was a hospital based cross sectional study, conducted over a period of 12 months (April 2016- March 2017). Blood sample for Ant-HCV antibody detection was taken at time of follow-up visit in the subspeciality clinic. These samples were processed in central laboratory for hep-C antibody, serum ferritin and transaminases levels. Anti-HCV antibody detection was done by BI-DOT machine. HCV RNA PCR was done to access viral load in all positive cases.Results: A total of 300 patients were enrolled in the study. There were 219 (73%) males and 81 (27%) females. The mean age of the study group was 7.59±3.6 years (range 1.5-18years). At the time of our study 277 (92.4%) cases were on one or the other chelating agent whereas 23 (7.6%) cases were not taking any kind of chelation therapy. Out of 300 patients, 72(24%) cases tested positive for anti HCV antibody. Out of 72 patients only 36(12%) patients had detectable viral load in RNA PCR. Mean age of the HCV positive cases (9.58±3.28) years was higher as compared to HCV negative cases (6.98±3.54). Maximum HCV positivity 20/38 (52.6%) was seen in 12-18 year age group; followed by 33/76 (43.4%) in 9-12yr age group. Significant association was observed between advancing age and prevalence of hepatitis C in thalassemia major patients (p=0.002). The number of blood transfusions received by anti-HCV positive children (Avg. Transfusion 185±98.40 ml/kg/year) was significantly higher than that by anti-HCV negative patients (Avg. Transfusion 102.8±71.20) (p value&lt;0.001). Maximum HCV positive cases 33 (45.83%) had total transfusions &gt;200 in a year followed by 15 (20.83%) cases with 151-200 transfusions (p&lt;0.001).Conclusions: Despite ELISA screening of blood donors, our study demonstrated high (24%) prevalence of transfusion transmitted hepatitis-C virus in thalassemic children which increases with increasing number of transfusions, it also correlates with rising serum ferritin level and SGPT level.

Diagnostic Dilemmas in Hepatitis C Virus Infection for Hemodialysis Patients.

Hepatitis C virus (HCV) is a leading cause of chronic hepatitis in dialysis patients. The diagnosis of HCV infection in these patients is predominantly based on laboratory tests because of the specificity of the clinical course of the disease. The present prospective study aimed at determining very accurately the prevalence rate of HCV infection in patients on dialysis by simultaneously testing them for anti-HCV and for HCV RNA levels. For the present cross-sectional longitudinal study we recruited and followed up 93 patients from St George University Hospital Hemodialysis Unit between July 2013 and December 2014. All patients were tested for anti-HCV and HCV RNA. The anti-HCV negative patients were tested for anti-HCV and HCV RNA at least twice at intervals of 6 months or more (up to 12 months). Anti-HCV antibodies were identified using a third generation ELISA assay. Commercial kits for real-time polymerase chain reaction (RT-PCR) were used to detect HCV RNA in the plasma and mononuclear cells. Aminotransferase and gammaglutamyl transpeptidase levels were studied to find if liver inflammation was present. The total seroprevalence in 68 patients was 20.6% (14). Of these, 10 patients were viremic (HCV RNA+/anti-HCV+), and 4 patients (5.9%) had discordant results (anti-HCV+/HCV RNA-). Acute hepatitis was detected in one patient. Duration of dialysis in HCV viremic patients was longer than that in aviremic patients (p=0.005). The present study suggests that HCV infection in dialysis patients can be diagnosed more accurately if these patients are tested using two diagnostic methods - a serological test and a biomolecular assay. Further studies with larger sample size may prove the feasibility of such approach for all dialysis patients in this country.

The prevalence of autoantibody and its relationship with genotypes of hepatitis C virus in patients with chronic hepatitis C virus infection.

The prevalence of autoantibody in the patients with chronic hepatitis C infection, and the relationship between the autoantibodies and HCV genotypes were investigated in this study. One hundred and eight anti-HCV positive and 86 anti-HCV negative patients were included in the study. Anti-HCV were studied by enzyme immunassay (EIA). HCV RNA was determined by real time polymerase chain reaction (PCR) and HCV genotypes were determined by a reverse-line blot hybridization. Anti-nuclear antibodies (ANA), anti-smooth muscle antibodies (ASMA), Anti-mitochondrial antibodies (AMA), liver kidney microsomal antibodies (LKM) were detected by indirect immunofluorescence assay. Among patients, 13 (12.03%) of 108 were positive for at least one autoantibody. The positivity was not observed in control group. The most prevalent autoantibody in anti-HCV positive group was ANA. ANA was positive in six HCV patients with genotype 1. In HCV patients with genotype 1, the frequencies of ANA, ASMA, AMA and LKM1 were six, two, three and one, respectively. In HCV patients with genotype 2, ANA was positive one patient and ASMA, AMA and LKM1 were not detected in HCV patients with genotype 2. In conclusion, the autoantibodies in patients with chronic hepatitis C in the study were low as compared to those reported in previous studies.

Characteristics of Seropositive Hepatitis B and C Thalassemia Major Patients in South-East of Iran

Background: Hepatitis is a serious blood born infection in patients with β-thalassemia major (β-TM). There was no previous report on hepatitis prevalence in patients with β-TM in Zabol, Iran. Objectives: The current study aimed to evaluate characteristics of hepatitis in patients with ᵝ-TM visiting Imam Khomeini Hospital of Zabol, in Sistan and Baluchistan province of Iran. Patients and Methods: There were 90 patients with ᵝ-TM. General demographic and laboratory results were obtained by interviews and reviewing clinical and laboratory histories. Data for anti-HCV, HBs antigen and anti-HBs were extracted from latest archived records, and analyzed using SPSS ver. 19. Results: Males and females comprised 51% and 49% of the patients, respectively. Mean age of the patients was 14.8±7.4 years old. Anti-HCV and HBs antigen were detected in 10% and 3.3% of the patients, respectively. Significant difference was found between the mean age of anti-HCV positive (20.1 ± 3.6, 95% CI: 17.3 - 22.9) and negative (14.2 ± 7.5, 95% CI: 12.5 - 15.9) patients (P = 0.02). Mean ferritin value was measured 4702 ± 2743 ng/mL. A significant correlation was observed between serum ferritin level and age (r = 0.3, P = 0.01). Hepatic enzymes, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) had mean levels of 51.8 ± 32.8 IU/L and 58.8 ± 59 IU/L, respectively. No significant association was identified between hepatitis status and either ferritin or hepatic enzymes levels. Conclusions: Results demonstrated the relatively high prevalence of HCV infection in the patients. Applying sensitive methods to screen blood units is recommended to minimize the risk of transfusion associated hepatitis. Keywords: β-Thalassemia Major, Anti-HCV, HBs Antigen, Anti-HBs, Ferritin

Hepatitis C Virus (HCV) Infection among Seronegative Patients undergoing Haemodialysis in a Remotely Located Tertiary Care Hospital of Northern India: Value of HCV-RNA and Genotypes.

Haemodialysis (HD) patients are at an increased risk of Hepatitis C virus (HCV) infection, which is significantly associated with increased morbidity and mortality. The aim of this study was to find the prevalence of HCV infection in anti-HCV antibody negative haemodialysis patients by Real-time PCR (RT-PCR) and value of HCV-RNA among seronegative patients undergoing haemodialysis in a remotely located tertiary care hospital. A total of 100 chronic renal failure patients on haemodialysis were studied. All the patients were screened for anti-HCV antibodies by ELISA test and for HCV-RNA by RT-PCR. The overall prevalence of HCV infection was 32%. Antibody positivity was 30% and HCV-RNA by RT-PCR was detected in 20%. HCV-RNA in seronegative patients was detected in 2.8%. Serological assays (30%) are quite reliable for detecting HCV infection in patients undergoing haemodialysis in our tertiary care hospital. Only a small proportion of them (2.8%) require the documentation of viral genome for current infection.

Occult hepatitis C virus infection in Iranian hemodialysis patients.

Background: Occult hepatitis C virus (HCV) infection is defined as the presence of HCV-RNA in liver or peripheral blood mononuclear cells (PBMCs) in the absence of detectable hepatitis C antibody (anti-HCV) or HCV-RNA in the serum. Low concentrations of HCV-RNA may be detected in PBMCs of hemodialysis (HD) patients and this could have a great impact on the management of HD patients. Objectives: The aim of this study was to detect the occult HCV infection in Iranian HD patients. Patients and Methods: A total of 70 anti-HCV negative HD patients from three dialysis units in Tehran, Iran were included in this study. In these cases, presence of HCV-RNA in plasma samples was tested by reverse transcriptase-nested polymerase chain reaction (RT-nested PCR). In cases with negative anti-HCV and plasma HCV-RNA, genomic HCV-RNA was checked in PBMC specimens by RT-nested PCR. Results: Seventy anti-HCV negative HD patients were enrolled in the study. 32.85% and 1.43% of cases had elevated levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) respectively. 7.14% of patients had elevated levels of both ALT and AST. HCV-RNA was negative in plasma samples of all anti-HCV negative HD subjects. The genomic HCV-RNA was not detected in any PBMC samples of HD cases with negative anti-HCV and plasma HCV-RNA. Conclusions: Occult HCV infection was not detected in our HD patients despite of elevated levels of liver enzymes in some participants. Further studies involving larger number of HD patients are required to elucidate the rate of occult HCV infection in HD cases.

Patients Screened by Two Methods; HCV Core Antigen and Polymerase Chain Reaction

BackgroundEnd-stage renal disease patients on chronic hemodialysis are among high risk groups for hepatitis C virus (HCV) infection for whom routine HCV screening is recommended. Anti-HCV antibody (ab) testing may not be reliable to detect all infected cases because of the blunted ab response due to depressed immune state in these patients. Using a more reliable, cost-effective and non-complex HCV screening test may be necessary in this group of patients for case finding and management, and also for prevention of infection spread.ObjectivesThe aim of this study was to find the prevalence of HCV infection in HCV ab negative hemodialysis patients by Real time PCR and total HCV core antigen (ag) test and comparing the results of the two tests.Patients and MethodsFrom a single hemodialysis center, 181 anti- HCV ab negative patients were screened by total HCV core ag using an ELISA kit. Real time PCR was used for determination of the virus and viral load quantity.ResultsAmong the 181 anti-HCV ab negative patients, 13 (7.2%) were positive for HCV core ag and 11 (6%) had detectable HCV RNA with a range of 40-336543 IU/ml by PCR. The two tests had a high measurement agreement (Kappa=0.82, P<0.001). Of the 13 patients with positive HCV core ag test results, 3 were negative for HCV RNA. Considering real time PCR for HCV RNA as the gold standard for HCV infection determination in this patient population, HCV core ag assay yielded a sensitivity of 90.9%, specificity of 98.2%, positive predictive value of 76.9% and negative predictive value of 99.4%.DiscussionThe rate of HCV infection among HCV ab negative hemodialysis patients was high. HCV core ag testing could be used as a sensitive method for HCV infection screening in this group of patients.

Prevalence of Occult Hepatitis B Virus Infection in Hemodialysis Patients

The prevalence of occult HBV infection depends on the prevalence of HBV infection in the general population. Hemodialysis patients are at increased risk for HBV infection. The aim of this study was to determine the prevalence of occult HBV infection in hemodialysis patients. Total of 98 patients undergoing hemodialysis in CHA Bundang Medical Center (Seongnam, Korea) were included. Liver function tests and analysis of HBsAg, anti-HBs, anti-HBc and anti-HCV were performed. HBV DNA testing was conducted by using two specific quantitative methods. HBsAg was detected in 4 of 98 patients (4.1%), and they were excluded. Among 94 patients with HBsAg negative and anti-HCV negative, one (1.1%) patient with the TaqMan PCR test and 3 (3.2%) patients with the COBAS Amplicor HBV test were positive for HBV DNA. One patient was positive in both methods. Two patients were positive for both anti-HBs and anti-HBc and one patient was negative for both anti-HBs and anti-HBc. The present study showed the prevalence of occult HBV infection in HBsAg negative and anti-HCV negative patients on hemodialysis at our center was 3.2%. Because there is possibility of HBV transmission in HBsAg negative patients on hemodialysis, more attention should be given to prevent HBV transmission.

Hepatitis C Virus Antibodies in Patients with Hemoglobinopathies - Decreased Prevalence but Continued Transfusion-Related New Cases in Two New York City Hospitals

Purpose: There are no recent studies on hepatitis C virus (HCV) in patients with hemoglobinopathies who are also at risk for liver damage from iron overload after multiple blood transfusions. The aims of this study were to determine prevalence and incidence of HCV antibodies (anti-HCV) in patients with sickle cell diseases, and to assess how anti-HCV status impacts hospital attendance. Methods: Medical records of patients with sickle cell diseases who tested positive for anti-HCV and patients screened for anti-HCV after January 2009 in 2 New York City hospitals (Interfaith Medical Center and Kings County Hospital) were reviewed. Of 189 patients, there were 130 S/S, 36 S/C, 15 S/beta+ thalassemia, 6 S/beta0 thalassemia, 1 C/C and 1 A/A with 76% fetal hemoglobin. Transfusion history, drug use, liver function tests, HCV RNA status and hospital attendances on the year of the latest negative anti-HCV test and 2 years following the first documented positive anti-HCV test were recorded. Results: Anti-HCV was detected in 26/189 patients (13.8%). HCV RNA was detected in 10 of the 26 cases, negative in 7 cases and unknown in 9 cases. In 4 of the 26 anti-HCV positive patients, we were able to establish that anti-HCV had been non reactive before year 2000. HCV RNA was detected in one of these 4 patients, negative in the second and unknown in two cases. These four patients had no risk factors other than recent polytransfusion. None of the 23/189 patients who never received blood transfusion tested positive for anti-HCV. There was a significant difference in ALT (but not AST) between anti-HCV positive and anti-HCV negative patients (ALT 76.90± 155.80 IU/L and 59.06± 298.48 IU/L, respectively; Wilcoxon, p=0.01). The annual hospital attendance for anti-HCV positive patients was unchanged at 2 year follow-up, and there was no difference compared to anti-HCV negative patients (3.5 attendances for each group, p=0.78). Conclusion: The prevalence of anti-HCV in patients with hemoglobinopathies has decreased from 16.5% in 2002 to 13.8% presumably due to systematic screening of blood products (Richard S, Billett HH, 2002). New reactive anti-HCV cases with detectable HCV RNA occur in patients requiring a high number of transfusions and suggest that anti-HCV is not 100% sensitive in diagnosing HCV in blood donors and a more sensitive assay may be warranted. Although anti-HCV positive status does not appear to worsen the course of sickle cell disease, we suggest periodic testing and education of patients who test positive for anti-HCV on the importance of their assessment by a hepatologist.

High prevalence of hepatitis C virus-ribonucleic acid positivity in anti-hepatitis C virus negative renal transplant patients

BackgroundHepatitis C virus (HCV) infection is common in renal transplant (RT) patients. Some of these patients remain anti-HCV negative despite presence of infection and these are identified by a positive HCV-ribonucleic acid (RNA) test. MethodsWe studied 404 RT patients for prevalence of HCV-RNA positivity in anti-HCV negative patients. Serum was tested for presence of anti-HCV antibodies using a third generation HCV micro-ELISA (enzyme-linked immunosorbent assay) test, which utilises a combination of HCV structural and nonstructural antigens. The RNA was extracted from patient serum for HCV viral quantification using Quiagen Ql Amp Viral RNA mini extraction kit. The HCV-RNA viral load was performed on Corbet Rotor Gene 3000 thermocycler using Taqman principle. ResultsAbout 308 patients were anti-HCV negative and 96 were anti-HCV positive, resulting in prevalence of overt HCV infection of 23.7%. A total of 130 anti-HCV negative patients tested positive for HCV-RNA making a prevalence of occult HCV infection of 42.2%. There was no significant difference in the rate of overt or occult HCV infection between males and females. Patients with HCV infection (whether overt or occult) had received more number of dialysis sessions (62.5 vs 32.2) and blood transfusions (2.78 vs 1.99) when compared to those without HCV infection (P=0.001). The mean duration on dialysis was also longer (8.15 months vs 4.53 months) in patients with HCV infection (P= 0.0001). ConclusionA direct test for HCV viraemia is important to accurately determine the epidemiology of HCV infection in RT patients who remain anti-HCV negative despite harbouring active HCV infection.

Characterization and Application of Monoclonal Antibody Against Hepatitis C Virus Nonstructual Protein Three

Developing assays for detecting HCV antigens could be beneficial because viral proteins appear earlier than antibodies and are more stable than RNA in the serum. Monoclonal antibody was prepared by immunization and cell fusion. Subclass, specificity, and efficiency of monoclonal antibodies were determined by ELISA. Epitope specificity of monoclonal antibodies was analyzed by ELISA additivity test. HCV antigen in serum of hepatitis patients was examined by double monoclonal antibody sandwich ELISA. Five hybridoma cell lines were screened and named HCV(1), HCV(2), HCV(3), HCV(4), and HCV(5). These five monoclonal antibodies had high specificity and efficiency. The additivity test showed that HCV(2), HCV(4), and HCV(5) recognized different epitopes, which can be matched in ELISA. Of 173 anti-HCV positive patients, 37 (21.4%) were positive for HCV antigen. Of 1498 anti-HCV negative patients, 10 (0.67%) were positive for HCV antigen. Fifty normal controls were negative for HCV antigen. HCV antigen detection had moderate agreement and correlation with HCV RNA detection (kappa=0.577, p<0.01; r=0.59, p<0.01). This result indicates that the monoclonal antibody against HCV NS(3) may be a potential diagnostic reagent, which would provide a foundation for developing a sandwich ELISA of HCV antigen detection.

New perspectives in occult hepatitis C virus infection

Occult hepatitis C virus (HCV) infection, defined as the presence of HCV RNA in liver and in peripheral blood mononuclear cells (PBMCs) in the absence of detectable viral RNA in serum by standard assays, can be found in anti-HCV positive patients with normal serum levels of liver enzymes and in anti-HCV negative patients with persistently elevated liver enzymes of unknown etiology. Occult HCV infection is distributed worldwide and all HCV genotypes seem to be involved in this infection. Occult hepatitis C has been found not only in anti-HCV positive subjects with normal values of liver enzymes or in chronic hepatitis of unknown origin but also in several groups at risk for HCV infection such as hemodialysis patients or family members of patients with occult HCV. This occult infection has been reported also in healthy populations without evidence of liver disease. Occult HCV infection seems to be less aggressive than chronic hepatitis C although patients affected by occult HCV may develop liver cirrhosis and even hepatocellular carcinoma. Thus, anti-HCV negative patients with occult HCV may benefit from antiviral therapy with pegylated-interferon plus ribavirin. The persistence of very low levels of HCV RNA in serum and in PBMCs, along with the maintenance of specific T-cell responses against HCV-antigens observed during a long-term follow-up of patients with occult hepatitis C, indicate that occult HCV is a persistent infection that is not spontaneously eradicated. This is an updated report on diagnosis, epidemiology and clinical implications of occult HCV with special emphasis on anti-HCV negative cases.

Prevention of Hepatitis B Virus and Hepatitis C Virus Transmission in Hemodialysis Centers: Review of Current International Recommendations

Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections in hemodialysis (HD) patients are associated with adverse outcomes, especially after kidney transplantation. In the HD setting, cross-contamination to patients via environmental surfaces, supplies, equipment, multiple-dose medication vials and staff members is mainly responsible for both HBV and HCV transmission. The incidence and prevalence of HBV in HD centers have dropped markedly as a result of isolation strategy for HBsAg positive patients, the implementation of infection control measures and the introduction of HBV vaccine. The incidence and prevalence of HCV infection among HD patients remain higher than the corresponding general population. There is ongoing debate as to whether isolation of HCV infected patients is needed to combat high anti-HCV seroconversion rates. The current guidelines do not recommend isolation or the use of dedicated machines for HCV infected patients, and rely on strict adherence to infection control measures for the prevention of HCV transmission in the HD setting. Investigations of dialysis associated outbreaks of HCV infection indicate that transmission most likely occurs because of inadequate infection control practices. Routine screening of anti-HCV negative patients, with HCV-antibody testing, and monthly monitoring of ALT levels is recommended to monitor transmission within centers. Prevention of transmission of HBV and HCV in the HD setting warrants a multi-faceted approach. Not enough stress can be placed on the importance of adequate infection control practices for the prevention of both infections. Prevention of HBV transmission is augmented by correct implementation of isolation strategies and the universal vaccination of susceptible patients.

Diagnostic performance of an immunoassay for simultaneous detection of HCV core antigen and antibodies among haemodialysis patients

Nosocomial transmission of HCV is a concern in haemodialysis (HD) units worldwide. Diagnosis of HCV infection among dialysis patients is currently based on the detection of anti HCV antibodies by ELISA, and is confirmed by HCV RNA. The average window period between HCV infection and seroconversion with new generations of HCV antibody tests remains approximately 70 days with more prolonged period among dialysis patients. In this study we assessed the diagnostic performance of an immunoassay designed for simultaneous detection of anti HCV antibodies and core antigen in one step in comparison to qualitative RT-PCR and anti HCV antibodies detection test among Egyptian haemodialysis patients. The studied patients were 39 chronic renal failure patients on maintenance haemodialysis. The results obtained in the present study revealed HCV infection of 56.4%. Combined Ag/Ab test detected 3 out of the 4 anti-HCV negative viraemic patients who were in the window period. The sensitivity, specificity and accuracy of the test were higher than that of anti HCV antibodies detection test (95.45%, 94.1% and 94.87% versus 81.8%, 88.23% and 84.6%) and they were raised to 100% on combining its positivity with liver enzymes elevation results. Therefore, this simple combined Ag/Ab test can be applied for early detection of HCV infection during window period among HD patients as an alternative to HCV RNA detection.