Detection Of Hepatitis C Virus Infection Research Articles

CDC's New Hepatitis C Virus Testing Recommendations for Perinatally Exposed Infants and Children: A Step Towards Hepatitis C Elimination.

New U.S. Centers for Disease Control and Prevention (CDC) guidelines for hepatitis C virus (HCV) testing of perinatally exposed infants and children released in 2023 recommend a nucleic acid test (NAT) for detection of HCV ribonucleic acid (i.e., NAT for HCV RNA) at 2-6 months of age to facilitate early identification and linkage to care for children with perinatally acquired HCV infection. Untreated hepatitis C can lead to cirrhosis, liver cancer, and premature death and is caused by HCV, a blood-borne virus transmitted most often among adults through injection drug use in the United States. Perinatal exposure from a birth parent with HCV infection is the most frequent mode of HCV transmission among infants and children. New HCV infections have been increasing since 2010, with the highest rates of infection among people aged 20-39 years, leading to an increasing prevalence of HCV infection during pregnancy. In 2020, the CDC recommended one-time HCV screening for all adults aged 18 years and older and for all pregnant persons during each pregnancy. Detecting HCV infection during pregnancy is key for the identification of pregnant persons, linkage to care for postpartum treatment, and identification of infants with perinatal exposure for HCV testing. It was previously recommended that children who were exposed to HCV during pregnancy receive an antibody to HCV (anti-HCV) test at 18 months of age; however, most children were lost to follow-up before testing occurred, leaving children with perinatal infection undiagnosed. The new strategy of testing perinatally exposed children at age 2-6 months was found to be cost-effective in increasing the identification of infants who might develop chronic hepatitis C. This report describes the current perinatal HCV testing recommendations and how they advance national hepatitis C elimination efforts by improving the health of pregnant and postpartum people and their children.

Cascade of Care for Hepatitis C Virus Infection Among Young Adults Who Inject Drugs in a Rural County in New Mexico.

Treatment for hepatitis C virus (HCV) infection is highly effective; however, people who inject drugs (PWID), the population most affected by HCV, may encounter barriers to treatment. We examined the cascade of care for HCV infection among young adult PWID in northern New Mexico, to help identify gaps and opportunities for HCV treatment intervention. Young adults (aged 18-29 y) who self-reported injection drug use in the past 90 days were tested for HCV antibodies (anti-HCV) and HCV RNA. We asked participants with detectable RNA to participate in an HCV education session, prior to a referral to a local health care provider for treatment follow-up, and to return for follow-up HCV testing quarterly for 1 year. We measured the cascade of care milestones ranging from the start of screening to achievement of sustained virologic response (SVR). Among 238 participants, the median age was 26 years and 133 (55.9%) were men. Most (90.3%) identified as Hispanic. Of 109 RNA-positive participants included in the cascade of care assessment, 84 (77.1%) received their results, 82 (75.2%) participated in the HCV education session, 61 (56.0%) were linked to care through a medical appointment, 27 (24.8%) attended the HCV treatment appointment, 13 (11.9%) attended their follow-up appointment, 6 (5.5%) initiated treatment, 3 (2.8%) completed treatment, and 1 (0.9%) achieved SVR. We observed a steeply declining level of engagement at each milestone step of the cascade of care after detection of HCV infection, resulting in a suboptimal level of HCV treatment and cure. Programs that can streamline testing and expand access to treatment from trusted health care providers are needed to improve the engagement of PWID in HCV treatment.

Expression of HCV genotype-4 core antigen in prokaryotic E. coli system for diagnosis of HCV infection in Egypt

BackgroundEgypt has a high prevalence of hepatitis C virus (HCV) infection with 92.5% of genotype-4. AimThis study aimed to clone and express the core gene of HCV genotype-4 for using it to develop a highly sensitive, specific, and cost-effective diagnostic assay for detecting HCV infection. MethodsUsing synthetic HCV genotype-4 core gene, pET15b as E. coli expression vector, and 1 mM lactose as inducer, the HCV core protein (MW 17 kDa) was expressed in the form of inclusion bodies (IBs) that was purified and solubilized using 8 M guanidinium HCl. The recombinant core protein was in vitro refolded by a rapid dilution method for further purification using weak cation exchange liquid chromatography. The immunogenicity of the purified protein was tested by ELISA using 129 serum samples. ResultsThe recombinant core protein was successfully expressed and purified. The results also showed that the in-house anti-HCV core assay is accurate, specific (~96.6%), and highly sensitive (~100%) in accordance with the commercial ELISA kit. ConclusionThe sensitivity, specificity, and reproducibility of the developed assay were high and promising to be used as a screening assay for detecting HCV infection.

Diagnostic Accuracy of Serum Hyaluronan for Detecting HCV Infection and Liver Fibrosis in Asymptomatic Blood Donors.

Background: The disease caused by hepatitis C virus (HCV) is asymptomatic, silent, and progressive liver disease. In HCV-infected patients the increase in serum HA is associated with the development of hepatic fibrosis and disease progression. Methods: HCV-RNA detection was performed in all serological samples of blood donors that tested positive using HCV Ultra ELISA. Determination of hyaluronan (HA) was performed in positive HCV samples using ELISA-like fluorometric method. The HA content was compared to HCV viral load, genotype of the virus, liver fibrosis as well as ALT and GGT liver biomarkers. Results: Persistently normal ALT (<40 U/L) and GGT (<50 U/L) serum levels were detected in 75% and 69% of the HCV-Infected blood donors, respectively. Based on ROC analysis, the HA value < 34.2 ng/mL is an optimal cut-off point to exclude HCV viremia (specificity = 91%, NPV = 99%). Applying HA value ≥34.2 ng/mL significant liver fibrosis (≥F2) can be estimated in 46% of the HCV-infected blood donors. HA serum level (≥34.2 ng/mL) associated with a high ALT level (>40 U/mL) can correctly identify HCV infection and probable liver fibrosis (sensitivity = 96% and specificity = 90%) in asymptomatic blood donors. Conclusions: A high level of HA (≥34.2 ng/mL) in association with ALT (≥40 U/L) in serum can provide a good clinical opportunity to detect HCV-infected asymptomatic persons that potentially require a liver biopsy confirmation and antiviral treatment to prevent the development of advanced liver fibrosis or cirrhosis.

The Biochemical investigation and analysis to determine a potent biomarker for HCC and HCV infections

The communicable Hepatitis C Virus (HCV) infection causes global health problems and deaths due to transmission by blood and body fluids and general symptoms include loss of appetite, fatigue, fever, abdominal pain, swelling, jaundice, and dark urine.This study contains a detailed study about the potent biochemical biomarkers for the diagnosis and detection of HCV infection. It includes the biochemical profiling relating to the HCV infection among both the genders of the human population is included in this paper.96 (5ml) EDTA blood samples of suspected HCV infected patients were collected from the different hospitals in Dehradun, Uttarakhand, India. The RNA extraction was performedwith the help of QIAamp Viral RNA mini kit (Cat. no. 52904). The HCV RNA quantification was achieved using the machine named Rotor Gene Q5 Plex Real-Time PCR and master mix was prepared by the Artus Amplification Kit from Qiagen.Out of 96 samples collected for HCV RNA biochemical profiling, 58 cases i.e., were found with the heavy load of virus and the study had 38 cases in which target virus was not detected. The biochemical investigation was performed on 96 cases with a high viral load of more than 34 IU/ml (≥34 IU/ml). The cases belonging to age 21-40 and 41-60 years were found to be most prominent among 35 and 37 cases respectively due to the repeated exposure to infected blood and the age group of 0-20 years was found to be least susceptible due to the protective antibody titer against HCV due to the vaccination in early age.

Hepatitis C virus infection in children and adolescents: A recent update

Hepatitis C virus (HCV) infection is an emerging problem for children and adolescents. The estimated 3.5–5 million children have chronic HCV infection globally. Early detection of HCV infection and early administration of appropriate treatment in childhood are necessary to prevent decompensated liver disease and hepatocellular carcinoma as in adults. Perinatal transmission is the most common route of HCV infection in children. An antibody-based test (anti-HCV) should be done in children at or after 18 months of age to detect HCV infection. Children who are anti-HCV-positive after 18 months of age should be further confirmed with an HCV-RNA assay after age 3 to recognize chronic hepatitis C infection. These patients should be further tested to determine the genotype. Children who are anti-HCV- and HCV-RNA-positive need to be treated with direct-acting antiviral. The purpose of the article is to discuss epidemiology, natural history, diagnosis, and management of HCV infection in children and adolescents.

Factors Associated With Testing for Hepatitis C Infections Among a Commercially Insured Population of Persons With HIV, United States 2008-2016.

Hepatitis C virus (HCV) infection is an important public health problem among people with HIV. People with HIV who are coinfected with HCV infection are at increased risk for cirrhosis, liver failure, and hepatitis C-related mortality; as such, national guidelines recommend that persons with HIV be tested for HCV infection. Data from the 2003-2017 IBM Watson Health MarketScan database were used for this study. We used diagnostic, procedural, and drug codes to identify patients with ≥1 inpatient or outpatient medical claim of HIV diagnosis. Patients with prior HIV or hepatitis C diagnoses were excluded. We calculated hepatitis C testing rates among newly diagnosed HIV-infected persons within 12 months of the initial HIV diagnosis date (January 1, 2008-December 31, 2016). We used Poisson regression to identify the factors associated with hepatitis C testing. Lastly, we assessed hepatitis C testing trends using the Cochran-Armitage test. The prevalence of testing for hepatitis C in newly identified persons with HIV (n = 46 277) was 50% within 12 months of the index HIV diagnosis. From 2008 to 2017, the testing rate increased by 13%. Significant predictors of hepatitis C testing were age, sex, and urbanicity. Women with HIV were less likely to have been tested compared with men (relative risk, 0.79; 95% CI, 0.77-0.81). Only 40% of patients between 50 and 59 years of age were tested for hepatitis C within 12 months of the index HIV diagnosis, while 56% of persons with HIV aged 20-29 years were tested for hepatitis C. Overall, 50% of newly diagnosed HIV patients were tested for hepatitis C within 12 months of HIV diagnosis. Although there were increases in hepatitis C testing rates over the study period, there were missed opportunities to detect HCV infection among people newly diagnosed with HIV.

High Prevalence of Hepatitis C Infection Among Adult Patients at Four Urban Emergency Departments - Birmingham, Oakland, Baltimore, and Boston, 2015-2017.

Identifying persons with hepatitis C virus (HCV) infection has become an urgent public health challenge because of increasing HCV-related morbidity and mortality, low rates of awareness among infected persons, and the advent of curative therapies (1). Since 2012, CDC has recommended testing of all persons born during 1945-1965 (baby boomers) for identification of chronic HCV infection (1); urban emergency departments (EDs) are well positioned venues for detecting HCV infection among these persons. The United States has witnessed an unprecedented opioid overdose epidemic since 2013 that derives primarily from commonly injected illicit opioids (e.g., heroin and fentanyl) (2). This injection drug use behavior has led to an increase in HCV infections among persons who inject drugs and heightened concern about increases in human immunodeficiency virus (HIV) and HCV infection within communities disproportionately affected by the opioid crisis (3,4). However, targeted strategies for identifying HCV infection among persons who inject drugs is challenging (5,6). During 2015-2016, EDs at the University of Alabama at Birmingham; Highland Hospital, Oakland, California; Johns Hopkins Hospital, Baltimore, Maryland; and Boston University Medical Center, Massachusetts, adopted opt-out (i.e., patients can implicitly accept or explicitly decline testing), universal hepatitis C screening for all adult patients. ED staff members offered HCV antibody (anti-HCV) screening to patients who were unaware of their status.* During similar observation periods at each site, ED staff members tested 14,252 patients and identified an overall 9.2% prevalence of positive results for anti-HCV among the adult patient population. Among the 1945-1965 birth cohort, prevalence of positive results for anti-HCV (13.9%) was significantly higher among non-Hispanic blacks (blacks) (16.0%) than among non-Hispanic whites (whites) (12.2%) (p<0.001). Among persons born after 1965, overall prevalence of positive results for anti-HCV was 6.7% and was significantly higher among whites (15.3%) than among blacks (3.2%) (p<0.001). These findings highlight age-associated differences in racial/ethnic prevalences and the potential for ED venues and opt-out, universal testing strategies to improve HCV infection awareness and surveillance for hard-to-reach populations. This opt-out, universal testing approach is supported by new recommendations for hepatitis C screening at least once in a lifetime for all adults aged ≥18 years, except in settings where the prevalence of positive results for HCV infection is <0.1% (7).

English

Screening of Hepatitis C virus (HCV) infection by molecular test is not routinely performed for the care of human immunodeficiency virus (HIV) infected patients in most countries of Sub-Saharan Africa such as Benin. The aim of this study was to assess the extent of HCV infection in patients infected with HIV in Cotonou. This study was conducted from February to June 2017 on HIV infected patients from the National Reference Center for Research and Management of HIV infection in Cotonou. Blood samples were collected from patients to detect anti-HCV antibody and HCV viral load. A total of 205 patients were tested, out of which 67.3% were females. Seroprevalence of anti-HCV antibody was 7.8% and HCV viral load was detectable in 3.4% of cases with a median of 2.200.000 IU/mL (6.3 Log10). Three out of seven patients (42.8%) had negative HCV serology with positive HCV RNA detection. In conclusion, the prevalence of HCV infection among HIV infected patients is not negligible in Cotonou. Universal access to molecular tests is needed in the country to detect HCV infection in these patients. Keywords: HCV, HIV, prevalence.

Detection Of Hepatitis C Virus (Hcv) Infection And Its Genotype In Patients At Hepatology Outpatient Clinic, Dr Soetomo General Hospital, Surabaya.

Hepatitis C virus (HCV) is an RNA virus that can cause liver inflammation (hepatitis) and has the potential to become chronic and can progress to liver cirrhosis and hepatocellular carcinoma. Detection of HCV RNA infection, and genotype/subtype of HCV was performed on 70 blood sera of patients at the Hepatology Outpatient Clinic of Dr Soetomo General Hospital, Surabaya, Indonesia. Detection of HCV infection was carried out by Anti-HCV determination using enzyme immunoassay (EIA) technique, detection of HCV RNA by Reverse Transcription Polymerase Chain Reaction (RT-PCR) technique based on genome regions NS5b and 5’UTR, followed by electrophoresis with agarose gel. In positive PCR results, HCV genotype/subtype was determined by direct sequencing method using ABI 310 sequencer and sequencing results were analyzed by comparing the products with previously published HCV nucleotides. Sera were obtained from 41 (58.6%) male and 29 (41.4%) female patients. Anti-HCV was found positive in 17/70 (24.29%) patients and 16/17 (94.1%) was proved to contain HCV RNA when determined by RT-PCR technique. Patients with positive HCV RNA have the potential to transmit HCV infection. From the genotype/subtype analysis of sequencing results we obtained 2/16 (12,5%), 3/16 (18,75%) and 6/16 (37,5%), 1/16 (6,25%), 1/16 (6, 25%), 2/16 (12,5%), 1/16 (6,25%) HCV genotypes 1, 2, and HCV subtypes 1b, 1c, 2a, 3a, 3k respectively.Conclusion: In patients who went to the Hepatology Outpatient Clinic, Dr. Soetomo General Hospital Surabaya, we found positive Anti-HCV was 24,29%. In 94,1% of patients with positive Anti-HCV, HCV RNA was still detected and HCV genotype 1 with subtype 1b were still dominant HCV subtypes.

Evaluation of in vitro screening and diagnostic kits for hepatitis C virus infection

BackgroundTo detect infection by hepatitis C virus (HCV), a reliable kit with high sensitivity and specificity is indispensable. Detection kits for anti-HCV antibodies (anti-HCV) are used for screening, and quantification kits for HCV RNA and core antigen are used for definite diagnosis of HCV infection. ObjectivesWe evaluated the performance of these kits using International Standards and a regional reference panel with HCV negative and positive specimens. Study designIn vitro diagnostic kits (10 anti-HCV, two HCV RNA, and three HCV core antigen) were included. ResultsNearly all specimens in the regional reference panel were correctly identified by all anti-HCV detection kits (one false-positive was observed in one kit). Both HCV RNA quantification kits also correctly identified and quantified HCV RNA titers, without genotype-specific differences. Among the HCV core antigen kits, International Standard values were inconsistent. The sensitivities of these kits were insufficient to detect HCV in positive specimens in the regional reference panel. ConclusionsIn vitro diagnostic kits assessing anti-HCV and HCV RNA have sufficient sensitivities and specificities to screen and detect HCV infection. However, HCV core antigen quantification kits have some limitations in their sensitivities and consistencies for diagnosis of HCV infection. Quality control with International Standards and a regional reference panel is important to maintain the performances of diagnostic kits for HCV infection and to verify the clinical reliability of these kits.

Evaluation of efficacy of serological methods for detection of HCV infection in blood donors: A single centre experience.

Background and Objective:Blood transfusion is an essential and life-saving medical intervention. Despite multiple preventive measures transfusion-transmitted hepatitis C virus (HCV) infection continues to be a major healthcare issue in Pakistan. This study was conducted at National Institute of Blood Diseases & Bone Marrow Transplantation to evaluate the frequency of active HCV infection with or without co-infection in blood donors and also to determine comparative efficacy of Multisure HCV antibody assay (MHAA); a new serological device.Methods:A total of 14652 blood donors visiting National Institute of Blood Diseases & Bone Marrow Transplantation (NIBD) Blood Bank from January 2013 to July 2014 were enrolled and screened for a range of blood borne infections such as HBV, HCV, HIV, malaria and syphilis. The HCV was screened simultaneously by Abbot Architect anti-HCV assay (CLIA) and MHAA. The active HCV infection was confirmed by nucleic acid testing (NAT) in reactive donors. Later; for determination of comparative efficacy of MHAA; all NAT positive samples were further tested using Monolisa™, HCV blot 3.0, Anti-HCV plus V2 and Anti-HCV-MPBIO-EIA.Results:The HCV reactive sera were observed in 1.563% (226) donors. The NAT confirmed active HCV infection in 138 donors. Overall 27.84% of HCV positive donors exhibited co-infection either with HBV (2.57%), syphilis (22.78%). Triple infection was not observed in any donor. The efficacy of MHAA is comparable to all the serological tests with a sensitivity of about 96.89%.Conclusion:Active HCV infection was present in 0.94% donors. With a sensitivity of 96.89% (95% CI: 95.66-98.12) the multi-parametric device MHAA can effectively detect HCV infection in donors. Thus, it can be used in limited health care settings for HCV screening.

A Preliminary Prospective Study for Seroprevalance of Hepatitis B Positivity in Hospital-Based Population from Bundelkhand Region of Uttar Pradesh

Background and study aim: Tuberculosis (TB) and hepatitis C virus (HCV) infection have emerged as major public health problems in Egypt. We aimed to determine the prevalence and risk factors for the HCV infection among patients with TB in Sohag and Qena governorates. Patients and methods: A cross-sectional study was carried out at Sohag and Qena university hospitals. Hundred thirty five tuberculosis patients fulfilled the inclusion criteria. HCV infection was diagnosed in 21/135 (6.4%). Results: Goza smoking (P value 0.01 Odd's Ratio 3.75, 95% confidence interval 0.24-0.44), history of operation (P value 0.001 ORs 7.67, 95% CI 0.165-0.263), blood transfusion (P value 0.004 ORs 7.2, 95% CI 0.103-0.362), presence of tattoos (P value 0.03 ORs 3.4, 95% CI 0.168-0.338), extra pulmonary tuberculosis (P value 0.004 ORs 3.5, 95% CI 2.341-3.384), low serum albumin (P value 0.002 ORs 0.5, 95% CI 0.068-0.317) were the main risk factors associated with HCV infection. Conclusion: Universal screening for HCV infection in TB patients is highly recommended with an urgent need to detect HCV infection in high-risk groups to prevent future HCV transmission as well as morbidity and mortality associated with TB.

The Prevalence of HCV Infection in Hemodialysis Population and Compared ELISA and PCR Methods for Detecting of HCV Infection

Background: Hemodialysis is one of the renal replacement therapies in patients with end-stage renal failure. The current study aimed at identifying the prevalence of hepatitis C virus (HCV) infection in hemodialysis population, and comparing serological (enzyme-linked immunosorbent assay) and molecular (polymerase chain reaction) methods to detect HCV infection in North of Iran. Methods: Serum samples from 162 patients undergoing chronic hemodialysis were collected in 2 hemodialysis units of Sari city (North of Iran). HCV RNAs were isolated from samples using RIBO-prep nucleic acid extraction kit (AmpliSens®, Russia). Total RNAs were extracted from samples and real-time polymerase chain reaction (PCR) was performed using HCV-FRT PCR kit (AmpliSense, Russia) according to the manufacturer’s instructions. Results: In the study, 7 (4.3%) cases were HCV-Ab positive and 155 (95.7%) HCV-Ab negative. Additionally, 11 patients (6.8%) were HCV-PCR positive, while 151 (93.2%) were HCV-PCR negative. Among 11 HCV-PCR positive patients, 7 (63.6%) were HCV-Ab positive and 4 (36.4%) were HCV-ab negative. HCV-ab test was not positive in any of the HCV-PCR negative patients. Conclusions: The results showed that the specificity of HCV-RNA detection was significantly higher than that of the conventional HCV-ab test. The gold standard test to confirm HCV positive should be PCR method.

Hepatitis C seroprevalence in an at-risk population in the southwest Madrid region of Spain

Abstract Introduction The estimated seroprevalence of hepatitis C virus (HCV) in Spain is 1.7%, but is much higher in the at-risk population. The most efficient national screening strategy is unclear. Aims To estimate the prevalence of HCV among the at-risk population seen in primary care (PC), and to determine their epidemiological profile. Materials and methods Cross-sectional descriptive prevalence study that included adult patients with risk factors for HCV infection seen in PC in the southwest Madrid region between 2010 and 2012. Results A total of 158 patients (men = 51.3%), mean age 46 years (SD = 16.6), were included. The most common risk factors were hypertransaminasaemia (44.3%) and major surgery (13.3%). Immigration, unsafe sexual practices, and tattoos or body piercing were more prevalent in patients younger than 45 years of age. Fifteen patients (9.5%) were positive for anti-HCV; 9 of these (5.7%) were HCV-ARN positive. Of the positive patients, 4 (44.4%) had significant fibrosis at diagnosis (F3–F4). Male patients had a higher rate of positive anti-HCV results (13.8 vs. 5.3%; p = 0.072), as did patients older than 45 years of age (12.8 vs. 6.3%; p = 0.167). Intravenous and intranasal drug use were associated with a higher rate of positive anti-HCV results (50 vs. 8.5%; p = 0.005 and 66.7 vs. 8.4%; p = 0.001, respectively). Conclusions Patients with risk factors for HCV infection have high seroprevalence. Screening programmes must therefore be implemented to detect HCV infection in this population in PC.

Seroprevalencia de hepatitis C en población con factores de riesgo del suroeste de la Comunidad de Madrid

IntroductionThe estimated seroprevalence of hepatitis C virus (HCV) in Spain is 1.7%, but is much higher in the at-risk population. The most efficient national screening strategy is unclear. AIMSTo estimate the prevalence of HCV among the at-risk population seen in primary care (PC), and to determine their epidemiological profile. Materials and methodsCross-sectional descriptive prevalence study that included adult patients with risk factors for HCV infection seen in PC in the southwest Madrid region between 2010 and 2012. ResultsA total of 158 patients (men=51.3%), mean age 46 years (SD=16.6), were included. The most common risk factors were hypertransaminasaemia (44.3%) and major surgery (13.3%). Immigration, unsafe sexual practices, and tattoos or body piercing were more prevalent in patients younger than 45 years of age. Fifteen patients (9.5%) were positive for anti-HCV; 9 of these (5.7%) were HCV-ARN positive. Of the positive patients, 4 (44.4%) had significant fibrosis at diagnosis (F3-F4).Male patients had a higher rate of positive anti-HCV results (13.8 vs. 5.3%; P=.072), as did patients older than 45 years of age (12.8 vs. 6.3%; P=.167). Intravenous and intranasal drug use were associated with a higher rate of positive anti-HCV results (50 vs. 8.5%; P=.005 and 66.7 vs. 8.4%; P=.001, respectively). ConclusionsPatients with risk factors for HCV infection have high seroprevalence. Screening programmes must therefore be implemented to detect HCV infection in this population in PC.

Comparison of Elecsys Anti-HCV II Assay With Other HCV Screening Assays.

Early detection of hepatitis C virus (HCV) is an important step in preventing progression to cirrhosis and hepatocellular carcinoma. Serologic assays for anti-HCV antibody are valuable first-line tests in the screening and diagnosis of HCV infection. This study's aim was to evaluate the sensitivity and specificity of Elecsys Anti-HCV II assay for HCV screening. A total of 1,044 routine sera, 20 known HCV-positive samples, plus 54 preselected weakly positive samples were tested for anti-HCV with Elecsys Anti-HCV II assay, Elecsys Anti-HCV assays, InTec HCV enzymoimmunoassay (EIA), and Livzon Anti-HCV EIA. Interference test was assessed with additional 423 specimens without clinical evidence of HCV infection: preselected HCV weak reactive samples; dialysis samples; anti-HBc (antibody to HBV core antigen) (+), anti-Treponema pallidum (+), and anti-HIV (+) sera; and samples form autoimmune/alcoholic hepatitis or systemic Lupus erythematosus (SLE). Discrepant results were evaluated with recombinant immunoblot assay. The seroconversion panels were evaluated to assess how early each assay could detect HCV infection. The specificity (99.81%) of the Elecsys Anti-HCV II assay was less than that with the two EIA comparison methods. However, false-negative results were easily seen in the EIA assays. When serial bleeds of HCV panels were compared with the above-mentioned methods, the assay detected acute HCV infection only 3.5 days after a positive HCV-RNA nucleic acid test and earlier than the comparator assays. Sensitivities and specificities of the anti-HCV assays were sufficiently high for use in this study. The Elecsys Anti-HCV II assay is suitable for screening and reliable early detection of HCV infection.

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.

Isolation, Cloning and Co-expression of Hepatitis C Virus Envelope Proteins: As Potential HCV Detecting Antigens

Aim: Approximately 3% of the world population is infected with Hepatitis C virus (HCV) which is the main cause of chronic liver disease. Blood transfusion is thought to be the leading cause of global epidemic of HCV. The envelope proteins E1 and E2 are involved in the early stages of the virus life cycle. These proteins have a major role in binding to receptors on the cell surface, fusion and integration of the virus into the host cell. Considering the potency of E1 and E2 in the development of diagnostic methods, the aim of our present study was co-expression of recombinant envelope proteins in eukaryotic HEK293 (human embryonic kidney) cells. Methods: The viral genomic RNA was used for cDNA (complementary DNA) synthesis. Isolation Original Research Article Mohammadipour et al.; BJMMR, 9(11): 1-8, 2015; Article no.BJMMR.16720 2 of HCV envelope proteins coding fragment was performed using cDNA and specific primers. The target gene was cloned into pcDNA3.1 expression vector, and transfected into HEK293 cells, an expression host. Accuracy of the cloning and expression was confirmed using PCR and Western blot analysis. Results: The isolation and cloning of the gene fragment encoding the E1 and E2 proteins was successful. Co-expression of these proteins was confirmed using monoclonal antibodies specific for each protein. Conclusion: This study showed that HEK293 host cell is suitable for the expression of hepatitis C virus E1 and E2 coding gene. These proteins can be used in numerous virological studies and detection of HCV infection.

Current molecular methods for the detection of hepatitis C virus in high risk group population: A systematic review.

Hepatitis C virus (HCV) is an emerging infection worldwide and the numbers of persons infected are increasing every year. Poor blood transfusion methods along with unsafe injection practices are potential sources for the rapid spread of infection. Early detection of HCV is the need of the hour especially in high risk group population as these individuals are severely immunocompromised. Enzyme Immunoassays are the most common detection techniques but they provide no evidence of active viremia or identification of infected individuals in the antibody-negative phase and their efficacy is limited in individuals within high risk group population. Molecular virological techniques have an important role in detecting active infection with utmost specificity and sensitivity. Technologies for assessment of HCV antibody and RNA levels have improved remarkably, as well as our understanding of how to best use these tests in patient management. This review aims to give an overview of the different serological and molecular methods employed in detecting HCV infection used nowadays. Additionally, the review gives an insight in the new molecular techniques that are being developed to improve the detection techniques particularly in High Risk Group population who are severely immunocompromised.