Significant Improvement in Diagnosis of Hepatitis C Virus Infection by a One-Step Strategy in a Central Laboratory: an Optimal Tool for Hepatitis C Elimination?

ABSTRACT The remarkable effectivity of current antiviral therapies has led to consider the elimination of hepatitis C virus (HCV) infection. However, HCV infection is highly underdiagnosed; therefore, a global strategy for eliminating it requires improving the effectiveness of HCV diagnosis to identify hidden cases. In this study, we assessed the effectiveness of a protocol for HCV diagnosis based on viral load reflex testing of anti-HCV antibody-positive patients (known as one-step diagnosis) by analyzing all diagnostic tests performed by a central laboratory covering an area of 1.5 million inhabitants in Barcelona, Spain, before (83,786 cases) and after (45,935 cases) the implementation of the reflex testing protocol. After its implementation, the percentage of anti-HCV-positive patients with omitted HCV RNA determination remarkably decreased in most settings, particularly in drug treatment centers and primary care settings, where omitted HCV RNA analyses had absolute reductions of 76.4 and 20.2%, respectively. In these two settings, the percentage of HCV RNA-positive patients identified as a result of reflex testing accounted for 55 and 61% of all anti-HCV-positive patients. HCV RNA results were provided in a mean of 2 days. The presence of HCV RNA and age of ≥65 years were significantly associated with advanced fibrosis, assessed using the serological FIB-4 index (odds ratio [OR], 5.92; 95% confidence interval [CI], 3.4 to 10.4). The implementation of viral load reflex testing in a central laboratory is feasible and significantly increases the diagnostic effectiveness of HCV infections, while allowing the identification of underdiagnosed cases.

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Hepatitis C Virus (HCV) diagnosis, epidemiology and access to treatment in a UK cohort

10.1101/216937 ◽

2017 ◽

Author(s):

Emily Adland ◽

Gerald Jesuthasan ◽

Louise Downs ◽

Victoria Wharton ◽

Gemma Wilde ◽

...

Keyword(s):

Hepatitis C Virus ◽

Viral Load ◽

Hepatitis C ◽

Positive Predictive Value ◽

Predictive Value ◽

Clinical Care ◽

Virologic Response ◽

Hcv Infection ◽

Hcv Rna ◽

Direct Acting Antiviral

ABSTRACTBackgroundAs direct acting antiviral (DAA) therapy is progressively rolled out for patients with hepatitis C virus (HCV) infection, careful scrutiny of HCV epidemiology, diagnostic testing, and access to care is crucial to underpin improvements in delivery of treatment.MethodsWe performed a retrospective study of HCV infection in a UK teaching hospital to evaluate the performance of different diagnostic laboratory tests, to describe the population with active HCV infection, and to determine the proportion of these individuals who access clinical care.ResultsOver a total time period of 33 months between 2013 and 2016, we tested 38,510 individuals for HCV infection and confirmed a new diagnosis of active HCV infection (HCV-Ag+ and/or HCV RNA+) in 359 (positive rate 0.9%). Our in-house HCV-Ab test had a positive predictive value of 87% when compared to repeat HCV-Ab testing in a regional reference laboratory, highlighting the potential for false positives to arise based on a single round of antibody-based screening. Of those confirmed Ab-positive, 70% were HCV RNA positive. HCV-Ag screening performed well, with 100% positive predictive value compared to detection of HCV RNA. There was a strong correlation between quantitative HCV-Ag and HCV RNA viral load (p<0.0001). Among the 359 cases of infection, the median age was 37 years, 85% were male, and 36% were in prison. Among 250 infections for which genotype was available, HCV genotype-1 (n=110) and genotype-3 (n=111) accounted for the majority. 117/359 (33%) attended a clinic appointment and 48 (13%) had curative treatment defined as sustained virologic response at 12 weeks (SVR12).ConclusionsHCV-Ab tests should be interpreted with caution as an indicator of population prevalence of HCV infection, both as a result of the detection of individuals who have cleared infection and due to false positive test results. We demonstrate that active HCV infection is over-represented among men and in the prison population. A minority of patients with a diagnosis of HCV infection access clinical care and therapy; enhanced efforts are required to target diagnosis and providing linkage to clinical care within high risk populations.ABBREVIATIONSDAADirect Acting AntiviralELISAEnzyme linked immunosorbent assayHCVHepatitis C VirusHCV-AbIgG antibody to Hepatitis C virusHCV-AgHepatitis C virus core antigenHCV RNAHepatitis C ribonucleic acid (viral load)MSMmen who have sex with menNATnucleic acid testingPCRpolymerase chain reaction (test for viral load)PPVpositive predictive valuePWIDpeople who inject drugsSDGSustainable Development GoalsSVRsustained virologic responseWHOWorld Health Organisation

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Higher levels of hepatitis C virus RNA found in blood donors co-infected with HIV as compared to HCV mono-infected donors

The Journal of Infection in Developing Countries ◽

10.3855/jidc.4767 ◽

2014 ◽

Vol 8 (08) ◽

pp. 1068-1071 ◽

Cited By ~ 4

Author(s):

Enagnon Kazali Alidjinou ◽

Donatien Moukassa ◽

Eben Ebatetou-Ataboho ◽

Gael Honal Mahoungou ◽

Jean-Paul Pambou ◽

...

Keyword(s):

Hepatitis C Virus ◽

Viral Load ◽

Hepatitis C ◽

Blood Donors ◽

Hiv Infections ◽

High Viral Load ◽

Hcv Infection ◽

Hcv Rna ◽

Infected People ◽

Sub Saharan

Introduction: Hepatitis C virus (HCV) infection and human immunodeficiency virus (HIV) infections are public health problems in sub-Saharan countries such as the Republic of Congo. HIV infection could impact the characteristics of HCV infection in co-infected people. We investigated HCV-HIV co-infection among blood donors in Congo. Methodology: Ninety-nine HIV-positive and/or HCV-seropositive blood donors were selected during screening and subsequently tested for aminotransferases and HCV RNA. Results: A total of 29 donors were found positive for HCV RNA (HCV-infected individuals), including 19/60 (31.66%) HIV donors (co-infected) and 10/39 (25.64%) non-HIV donors (mono-infected). Most of the co-infected donors (17/19) displayed a high viral load (> 5 log). The median HCV RNA level was at least 2 logs higher in co-infected people. The levels of alanine aminotransferase (ALT) were also slightly higher in co-infected donors than in HCV mono-infected donors. Conclusion: This study reports HCV-HIV co-infection among blood donors in Congo and shows that HCV viral load is higher in HIV donors.

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Prevalence and Risk Factors of Hepatitis C Virus (HCV) in Tehsil Batkhela District Malakand, KPK, Pakistan

International Journal of Contemporary Research and Review ◽

10.15520/ijcrr/2018/9/06/525 ◽

2018 ◽

Vol 9 (06) ◽

pp. 20251-20256

Author(s):

Mudassir Khan ◽

Shahrukh Khan ◽

Shohra Haider ◽

Fazal Jalil ◽

Muhsin Jamal ◽

...

Keyword(s):

Risk Factors ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Skin Color ◽

Significant Risk ◽

Rapid Test ◽

Prevalence Ratio ◽

Hcv Infection ◽

Hcv Rna ◽

Common Symptom

Background: Prevalence of Hepatitis C viral infection and its major risk factors has been found out in population of Batkhela, Khyber Pakhtunkhwa, Pakistan by taking number of volunteers from the interested area. HCV prevalence has not been researched in recent time here in this area, so that’s why we contributed. Materials and Methods: Ab rapid test cassette serum/plasma (USA) kit has been used for the mentioned purpose following by ELISA and finally PCR to find out active infection of virus. ICT positive individuals were reconfirmed by ELISA and then ELISA positive samples were carefully investigated by RT-PCR for Hepatitis C Virus. Results: The study population was of 770 volunteers belonging to the mentioned area of research, 453 males and 317 females. The overall prevalence was found to be 5.32% of HCV in Batkhela. This prevalence ratio was 3.12% in males and 2.20 % in females. 3rd generation ELISA was used to refine ICT positive samples which showed that 37 of the ICT positive samples had antibodies detected by ELISA. To find out active HCV infection, ELISA positive samples were refined by real time PCR which showed 2.98% of prevalence of active HCV infection in Batkhela based on HCV RNA in their blood. Principle Conclusion: Overall prevalence was found 5.32%, contaminated reused syringes and blades at Barbour’s shop, blood transfusion, surgical operations and unhygienic food in stalls etc were found significant risk factors for acquiring HCV infection. Body weakness and pale yellow skin color was common symptom in HCV positive volunteers. Safe sexual activities, blood screening before donation and sterilizing surgical equipment’s can protect us from Hepatitis C Virus.

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Hepatitis C Virus Prevalence and Risk Factors in a Village in Northeastern Romania—A Population-Based Screening—The First Step to Viral Micro-Elimination

Healthcare ◽

10.3390/healthcare9060651 ◽

2021 ◽

Vol 9 (6) ◽

pp. 651

Author(s):

Laura Huiban ◽

Carol Stanciu ◽

Cristina Maria Muzica ◽

Tudor Cuciureanu ◽

Stefan Chiriac ◽

...

Keyword(s):

Risk Factors ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Antiviral Treatment ◽

Population Based ◽

Hcv Infection ◽

World Health ◽

Hcv Rna ◽

Virus Eradication ◽

Tertiary Center

(1) Background: The World Health Organization adopted a strategy for the Global Health Sector on Viral Hepatitis in 2016, with the main objective of eliminating hepatitis C virus (HCV) by 2030. In this work, we aimed to evaluate the prevalence of HCV infection and risk factors in a Romanian village using population-based screening as part of the global C virus eradication program. (2) Methods: We conducted a prospective study from March 2019 to February 2020, based on a strategy as part of a project designed to educate, screen, treat and eliminate HCV infection in all adults in a village located in Northeastern Romania. (3) Results: In total, 3507 subjects were invited to be screened by rapid diagnostic orientation tests (RDOT). Overall, 2945 (84%) subjects were tested, out of whom 78 (2.64%) were found to have positive HCV antibodies and were scheduled for further evaluation in a tertiary center of gastroenterology/hepatology in order to be linked to care. In total, 66 (85%) subjects presented for evaluation and 55 (83%) had detectable HCV RNA. Of these, 54 (98%) completed antiviral treatment and 53 (99%) obtained a sustained virological response. (4) Conclusions: The elimination of hepatitis C worldwide has a higher chance of success if micro-elimination strategies based on mass screening are adopted.

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Prospective study of hepatitis C virus infection in hemodialysis patients by monthly analysis of HCV RNA and antibodies

Canadian Journal of Microbiology ◽

10.1139/w03-065 ◽

2003 ◽

Vol 49 (8) ◽

pp. 503-507 ◽

Cited By ~ 25

Author(s):

Regina Moreira ◽

João Renato Rebello Pinho ◽

Jorge Fares ◽

Isabel Takano Oba ◽

Maria Regina Cardoso ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Hemodialysis Patients ◽

Hcv Infection ◽

Hcv Rna ◽

Hepatitis C Virus Infection ◽

Serum Samples ◽

Hcv Genotypes ◽

High Prevalence ◽

Time Required

The aims of this study were to (i) evaluate the prevalence and the incidence of hepatitis C virus (HCV) infection in hemodialysis patients in two different centers in São Paulo (Brazil), (ii) determine the time required to detect HCV infection among these patients by serology or PCR, (iii) establish the importance of alanine aminotransferase determination as a marker of HCV infection, and (iv) identify the HCV genotypes in this population. Serum samples were collected monthly for 1 year from 281 patients admitted to hospital for hemodialysis. Out of 281 patients, 41 patients (14.6%) were HCV positive; six patients seroconverted during this study (incidence = 3.1/1000 person-month). In 1.8% (5/281) of cases, RNA was detected before the appearance of antibodies (up to 5 months), and in 1.1% (3/281) of cases, RNA was the unique marker of HCV infection. The genotypes found were 1a, 1b, 3a, and 4a. The presence of genotype 4a is noteworthy, since it is a rare genotype in Brazil. These data pointed out the high prevalence and incidence of HCV infection at hemodialysis centers in Brazil and showed that routine PCR is fundamental for improving the detection of HCV carriers among patients undergoing hemodialysis.Key words: HCV genotypes, hemodialysis, hepatitis C, PCR, prevalence, incidence.

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Hepatitis C Virus Clearance after Discontinuation of Pegylated Interferon Alpha-2a Monotherapy in a Child

Case Reports in Medicine ◽

10.1155/2012/597348 ◽

2012 ◽

Vol 2012 ◽

pp. 1-4

Author(s):

Takeshi Endo ◽

Koichi Ito ◽

Tokio Sugiura ◽

Kenji Goto

Keyword(s):

Hepatitis C Virus ◽

Viral Load ◽

Hepatitis C ◽

Antiviral Therapy ◽

Interferon Alpha ◽

Antiviral Treatment ◽

Pegylated Interferon ◽

Hcv Rna ◽

Pegylated Interferon Alpha ◽

Present Patient

The present patient was a 4-year-old boy. His hepatitis C virus genotype was 2a, and his viral load was high (1400,000 U/mL). The pretreatment liver biopsy revealed no fibrosis or malignancy and mild chronic hepatitis; his Knodell's histological activity (HAI) score was 4. Single nucleotide polymorphism of IL28B (rs8099917) was major type. The patient began antiviral treatment with pegylated interferon alpha 2a (90 μg/week). At week 9, serum HCV RNA became undetectable, with a sensitivity of 50 copies/mL. Antiviral treatment was discontinued at week 11 because the ALT level increased to 610 U/L. After discontinuation of therapy, the patient’s serum HCV RNA status became positive again. The serum viral load increased to 100,000 U/mL. During this period, he had been observed without medication. Sixteen months after stopping treatment, serum HCV became undetectable. Over a 4-year period, HCV RNA became negative and his anti-HCV antibody titer gradually decreased. In conclusion, though antiviral therapy resulted in failure or incomplete therapy, a reduced viral load resulted in viral clearance in the present patient. Interleukin 28B genotype might have association with the clearance of hepatitis C virus after discontinuation of antiviral therapy.

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Hepatitis C virus depends on E-cadherin as an entry factor and regulates its expression in epithelial-to-mesenchymal transition

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1602701113 ◽

2016 ◽

Vol 113 (27) ◽

pp. 7620-7625 ◽

Cited By ~ 35

Author(s):

Qisheng Li ◽

Catherine Sodroski ◽

Brianna Lowey ◽

Cameron J. Schweitzer ◽

Helen Cha ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Cell Surface ◽

Viral Entry ◽

Epithelial To Mesenchymal Transition ◽

Hcv Infection ◽

Hcv Rna ◽

Entry Site ◽

Mesenchymal Transition ◽

E Cadherin

Hepatitis C virus (HCV) enters the host cell through interactions with a cascade of cellular factors. Although significant progress has been made in understanding HCV entry, the precise mechanisms by which HCV exploits the receptor complex and host machinery to enter the cell remain unclear. This intricate process of viral entry likely depends on additional yet-to-be-defined cellular molecules. Recently, by applying integrative functional genomics approaches, we identified and interrogated distinct sets of host dependencies in the complete HCV life cycle. Viral entry assays using HCV pseudoparticles (HCVpps) of various genotypes uncovered multiple previously unappreciated host factors, including E-cadherin, that mediate HCV entry. E-cadherin silencing significantly inhibited HCV infection in Huh7.5.1 cells, HepG2/miR122/CD81 cells, and primary human hepatocytes at a postbinding entry step. Knockdown of E-cadherin, however, had no effect on HCV RNA replication or internal ribosomal entry site (IRES)-mediated translation. In addition, an E-cadherin monoclonal antibody effectively blocked HCV entry and infection in hepatocytes. Mechanistic studies demonstrated that E-cadherin is closely associated with claudin-1 (CLDN1) and occludin (OCLN) on the cell membrane. Depletion of E-cadherin drastically diminished the cell-surface distribution of these two tight junction proteins in various hepatic cell lines, indicating that E-cadherin plays an important regulatory role in CLDN1/OCLN localization on the cell surface. Furthermore, loss of E-cadherin expression in hepatocytes is associated with HCV-induced epithelial-to-mesenchymal transition (EMT), providing an important link between HCV infection and liver cancer. Our data indicate that a dynamic interplay among E-cadherin, tight junctions, and EMT exists and mediates an important function in HCV entry.

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Characterization of Occult Hepatitis C Virus (Hcv) Infection among Hemodialysis Patient

IIUM Medical Journal Malaysia ◽

10.31436/imjm.v16i1.1138 ◽

2017 ◽

Vol 16 (1) ◽

Author(s):

Siti Nurul Fazlin Abdul Rahman ◽

Hairul Aini binti Hamzah ◽

Mohammed Imad Mustafa ◽

Mohamed Hadzri Hasmoni

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Hemodialysis Patients ◽

Hcv Infection ◽

Viral Rna ◽

Cell Counting ◽

Hcv Rna ◽

Viral Rnas ◽

Occult Hepatitis ◽

Occult Hepatitis C

Introduction: The existence of new entity called occult hepatitis C virus (HCV) has become a raising and escalating concern among healthcare professionals worldwide. It is defined by the presence of viral RNA in liver and/or peripheral blood mononuclear cells (PBMCs) within non HCV-infected patients. Previous study had shown the occult HCV is infectious and capable of transmitting the virus to another host. Till today, HCV infection remains common among hemodialysis patients despite having the best preventive plans. Because of this, there is a significant concern about the source of viral transmission. The aim of the study was to identify and characterize occult HCV infection in PBMC sample of hemodialysis patients. This was an observational and cross sectional study. Materials and method: PBMCs were isolated from the whole blood using Ficoll-gradient centrifugation technique. The PBMCs were then subjected for cell counting and stored in -70O C until further used. HCV RNA were extracted from these cells and viral RNA were subjected for molecular assays, immune cells analysis and cells culture. Results: PBMCs were isolated from eleven (11) study patients and five (5) anti-HCV positive (control) patients. By using automated flow cytometry, PBMCs of each sample were counted and the average number of cells obtained range from 2x104 to 5x106 cells/ ml. Viral RNAs were extracted and quantitatively measured by using NanoDrop Spectrophotometers. The viral RNAs concentration obtained were between 24.7 and 258.9 ng/ml. The RNAs would be subjected for purification (ethanol precipitation) and further assays. Conclusion: The final findings might contribute to the clinical management of dialysis patients.

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Prevalence of Hepatitis C Virus in the Population of Albania for the Period 2007-2010

Open Access Macedonian Journal of Medical Sciences ◽

10.3889/oamjms.2014.094 ◽

2014 ◽

Vol 2 (3) ◽

pp. 525-528 ◽

Cited By ~ 1

Author(s):

Hysaj Vila Brunilda ◽

Shundi Lila ◽

Abazaj Erjona ◽

Bino Silva ◽

Rexha Tefta

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Age Groups ◽

Viral Disease ◽

Hcv Infection ◽

University Hospital ◽

P Value ◽

Hcv Rna ◽

Significant Difference ◽

Males And Females

BACKGROUND: Hepatitis C is a blood-borne, infectious, viral disease that is caused by a hepatotropic virus called Hepatitis C virus (HCV).AIM: The aim of this study is to determine the prevalence of active HCV infection (HCV–RNA) in the cases that were anti-HCV positive.MATERIAL AND METHODS: Plasma of 301 high-risk for HCV infection consecutive from University Hospital Centre “Mother Theresa” Tirana-Albania, during January 2007 to December 2010 was included in this study. To identify the presence of HCV RNA, the samples were examined by Cobas Amplicor HCV test (qualitative method).RESULTS: From 301 samples analyzed in total, 214 of them resulted positive for the presence of HCV-RNA's, corresponding to a prevalence of 71.1%, with 95% CI interval [65.8 - 75.9] for value of χ2 = 52.7 p value <0.0001. Divide by the sex 56% were males and 44% females, with statistically significant difference between them for value χ2 =4306 p value=0.0380. Among the age groups the highest prevalence was observed in the age groups > 25 years with a significant difference with other age groups for p value <0.001.CONCLUSION: Among tested samples, 71.1 % were confirmed to be positive for HCV –RNA infections. The prevalence of male was highest compared to female. For males and females infected the prevalence was highest in the age group of > 25 years.

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Robust Production of Infectious Hepatitis C Virus (HCV) from Stably HCV cDNA-Transfected Human Hepatoma Cells

Journal of Virology ◽

10.1128/jvi.79.22.13963-13973.2005 ◽

2005 ◽

Vol 79 (22) ◽

pp. 13963-13973 ◽

Cited By ~ 120

Author(s):

Zhaohui Cai ◽

Chen Zhang ◽

Kyung-Soo Chang ◽

Jieyun Jiang ◽

Byung-Chul Ahn ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Cell Lines ◽

Culture System ◽

Infectious Virus ◽

Hcv Infection ◽

Hcv Rna ◽

Human Hepatoma ◽

Stable Cell Lines ◽

Increased Risk

ABSTRACT Hepatitis C virus (HCV) chronically infects approximately 170 million people worldwide, with an increased risk of developing cirrhosis and hepatocellular carcinoma. The study of HCV replication and pathogenesis has been hampered by the lack of an efficient stable cell culture system and small-animal models of HCV infection and propagation. In an effort to develop a robust HCV infection system, we constructed stable human hepatoma cell lines that contain a chromosomally integrated genotype 2a HCV cDNA and constitutively produce infectious virus. Transcriptional expression of the full-length HCV RNA genome is under the control of a cellular Pol II polymerase promoter at the 5′ end and a hepatitis delta virus ribozyme at the 3′ end. The resulting HCV RNA was expressed and replicated efficiently, as shown by the presence of high levels of HCV proteins as well as both positive- and negative-strand RNAs in the stable Huh7 cell lines. Stable cell lines robustly produce HCV virions with up to 108 copies of HCV viral RNA per milliliter (ml) of the culture medium. Subsequent infection of naïve Huh7.5 cells with HCV released from the stable cell lines resulted in high levels of HCV proteins and RNAs. Additionally, HCV infection was inhibited by monoclonal antibodies specific to CD81 and the HCV envelope glycoproteins E1 and E2, and HCV replication was suppressed by alpha interferon. Collectively, these results demonstrate the establishment of a stable HCV culture system that robustly produces infectious virus, which will allow the study of each aspect of the entire HCV life cycle.

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