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Early diagnosis remains key for effective prevention and treatment. Unfortunately, current screening with anti-hepatitis C virus antibody (anti-HCV Ab) test may have limited utility in the diagnosis of HCV infection and reinfection. This is of special concern to at-risk population, such as immunocompromised hosts and end-stage renal failure patients on hemodialysis. HCV antigen (Ag) could be useful in identifying the ongoing infection in such clinical scenarios. Hence, we aimed to study the utility of HCV Ag testing for the diagnosis of acute and chronic hepatitis C. Of 89 samples studied, 19 were from acute hepatitis C patients who were immunocompromised or were on hemodialysis, 43 were from active chronic hepatitis C patients and 27 were from patients treated for chronic hepatitis C. All samples were tested for HCV Ag using the Abbott ARCHITECT HCV Ag assay. HCV Ag was reactive in 19/19 samples from acute hepatitis C patients and 42/43 samples from active chronic hepatitis C patients. It was nonreactive in all samples from treated patients. The test showed a sensitivity and specificity of 98.4% and 100.0%, respectively. The positive and negative predictive values were 100.0% and 96.4%, respectively. The HCV antigen test has high clinical sensitivity and specificity and is useful for the diagnosis of acute and chronic hepatitis C infection in at-risk and immunocompromised patients. Its short turnaround time and relatively low cost are advantageous for use in patients on hemodialysis and other at-risk patients who require monitoring of HCV infection and reinfection.

In developing nations, the major cause for liver infection that lead to liver fibrosis and cancer is the virus type C. The research has been conducted on 800 individuals assumed to be infected with hepatitis C virus, those individuals have been admitted to AL-Hilla Teaching Hospital ,Central Public Health laboratory, to the possible relation of serum interferon -α, CD56, IL-10 and IL-12 in case infection of virus type C comparison to normal healthy control groups. ELISA assay was used for detection antibodies to hepatitis C virus antigen as well as its immunological response in the subjects which were infected. The result indicated that just 40 from 800 individuals (12-62) years old had HCV positive which represent (5.2%) with high frequency in male( 67.3%/), than female(32.6%) groups . The study also show elevated of IL-10 level in all groups of patients compared to the groups of normal healthy controls (P0.05). The highest level was reached to 81.61pg/ml and 90.69pg/ml in male and female respectively. The IL-12 level also revealed a significant increase level in all groups of patients compared to the group of normal healthy controls (P0.05). The highest level for it was reached to 99.25 pg/ml and 125.33pg/ml in male and female respectively. The study also revealed enhancement of IFN- α levels in all groups of patients compared to the groups of normal healthy controls (P0.05) the higher level which was 201.69 pg/ml and 311.71pg/ml male and female respectively. The results also shows increase level of CD56 in all groups of patients compared to the group of normal healthy controls (P0.05). The highest level was reached to 27.984ng/ml and 30.190ng/ml in male and female respectively

AIMS: Hepatitis C virus antigen (HCV‐Ag) detection requires retesting for samples with grey zone results (GzR), adding cost and time and decreasing reliability. Our aim in this study was to evaluate the frequency and significance of GzR during the use of the automated Architect HCV‐Ag assay in routine clinical practice. METHODS AND RESULTS: We studied HCV‐Ag levels in 952 serum samples using the ARCHITECT HCV‐Ag assay. GzR were detected in 33 samples; 25 were reactive on retesting and 19 were anti‐HCV positive. Seventeen of these 19 samples were tested for HCV‐RNA and were all reactive (viral loads

UNLABELLED The current standard in diagnosing hepatitis C virus (HCV) infection requires two sequential steps: anti-HCV test to screen, followed by HCV RNA reverse-transcription polymerase chain reaction to confirm viremic HCV (V-HCV) infection. HCV core antigen tests provided potential for possible one-step diagnosis. However, low sensitivity and specificity limit their clinical utility. The present study developed a novel HCV antigens enzyme immunoassay (HCV-Ags EIA) and assessed its sensitivity, specificity, and utility for one-step diagnosis of V-HCV infection using 365 serum specimens, including 176 without and 189 with V-HCV infection. First, we confirmed the presence of HCV nonstructural proteins 3, 4b, and 5a besides HCV core antigen during HCV infection and developed a novel HCV-Ags EIA through simultaneous detection of all four HCV proteins. For the first time, the present study demonstrated that serum sample denaturation decreases the test specificity due to release of HCV-Ags sequestered in HCV immune complexes and should not be used in any HCV-Ags, including all the current HCV core antigen assays. On the other hand, using sample nondenaturation, the HCV-Ags EIA results showed 98.9% specificity and 100% sensitivity compared to serum anti-HCV and HCV RNA reverse-transcription polymerase chain reaction results. Using serum sample dilution, and nondenaturation, the lowest limits of detection of the HCV-Ags EIA were equivalent to serum HCV RNA levels of approximate 150-250 IU/mL. CONCLUSIONS The highly specific and sensitive HCV-Ags EIA developed in the present study has the lowest limit of detection equivalent to serum HCV RNA levels of 150-250 IU/mL; using nondenaturation of serum samples, our HCV-Ags EIA reliably differentiated V-HCV infection from resolved HCV infection, accomplishing screening and diagnosis of V-HCV infection in one step. (Hepatology 2016;64:415-424).

A method for detection and identification of the hepatitis C virus antigen (HCVcoreAg) in human serum with consideration for possible amino acid substitutions is proposed. The method is based on a combination of biospecific capturing and concentrating of the target protein on the surface of the chip for atomic force microscope (AFM chip) with subsequent protein identification by tandem mass spectrometric (MS/MS) analysis. Biospecific AFM-capturing of viral particles containing HCVcoreAg from serum samples was performed by use of AFM chips with monoclonal antibodies (anti-HCVcore) covalently immobilized on the surface. Biospecific complexes were registered and counted by AFM. Further MS/MS analysis allowed to reliably identify the HCVcoreAg in the complexes formed on the AFM chip surface. Analysis of MS/MS spectra, with the account taken of the possible polymorphisms in the amino acid sequence of the HCVcoreAg, enabled us to increase the number of identified peptides.

Обзор литературы посвящен возможностям применения в диагностике гепатита С (ГС) серологических тестов, выявляющих капсидный антиген вируса гепатита С (ВГС). Наиболее распространенные серологические тесты, используемые для первичной диагностики ВГС-инфекции, основаны на обнаружении антител к вирусу (анти-ВГС). Однако эти тесты не позволяют дифференцировать случаи перенесенной и активной инфекции. В то же время определение антигена ВГС позволяет выявлять активную ВГС-инфекцию. Интерес к данному тесту в последнее время возрастает в связи с увеличением потребности в эффективных высокопроизводительных тест-системах для проведения массового скрининга активной ВГС-инфекции, а также контроля эффективности антивирусной терапии. Рассмотрены существующие тесты для выявления антигена ВГС, опыт их применения и экономическая эффективность диагностики ГС., This review focuses on the use of serological tests that detect hepatitis C virus (HCV) antigen. The most common serological tests used for the primary diagnosis of HCV infection are based on the detection of antibodies to the virus (anti-HCV). However, these tests do not allow distinguish cases of resolved and active infection. Meanwhile, the definition of HCV antigen allows the identification of persons with active HCV infection. Interest in this test has recently grown due to an increase in the need for effective, high-performance and cheap tests for mass screening of active HCV infection, as well as monitoring the effectiveness of antiviral therapy. This review examines existing tests for the detection of HCV antigen, its application in clinical practice, and data on the cost-effectiveness of implementation of these tests into hepatitis C diagnostics.

Background: Of the donor corneas rejected for transplantation, the largest group is that from donors testing seropositive for hepatitis C virus (HCV). In situations of severe shortage in supply of donor corneal tissue, we may consider the use of seropositive donors for transplantation if we can prove with high certainty the absence of HCV RNA in the donor corneal tissue. Polymerase chain reaction (PCR) is a highly sensitive and specific technique for direct detection of HCV RNA and can be used for this purpose. Nevertheless, it is not applicable for routine clinical use in most eye departments due to its unavailability and cost effectiveness.Purpose: To study the possible use of immunohistochemical method for detection of HCV antigen in corneal tissue of seropositive donors and correlate the results with those of PCR. Immunohistochemical methods have not yet been studied in donor corneal tissue.Materials and methods: Eight corneas of 4 seropositive and 8 corneas of 4 seronegative corneal donors were studied by immunohistochemical and PCR methods for the presence of HCV antigen in their corneal tissue and sera.Results: HCV RNA was not detected in the sera and corneal tissue of all seropositive and seronegative corneal donors by either PCR and immunohistochemical methods.Conclusion: Although the study is too small for conclusive results, the correlation between the immunohistochemical and PCR studies for direct detection of HCV antigen in corneal tissue of seropositive donors may raise the possibility of using the immunohistochemical method for screening of donor corneas for the detection of HCV antigen. A larger prospective study investigating the sensitivity, specificity and clinical applicability of the immunohistochemical method is warranted.

SUMMARY. Reduction of the window period of hepatitis C virus (HCV) infection represents an important goal in the transfusional and diagnostic setting. A prototype assay designed to simultaneously detect circulating HCV antigen and anti-HCV, has been developed. Aim of this study was to evaluate the performance of this new assay in terms of specificity and sensitivity and to compare its efficacy with commercial assays. To evaluate the specificity of the assay, 400 samples from the general population and 100 difficult sera, negative for anti-HCV, were tested. To assess sensitivity, the new test was used on 76 PCR-positive and anti-HCV negative sera, seven natural or commercial seroconversion panels that included 17 RNA-positive and anti-HCV negative sera and 31 anti-HCV positive sera, 20 weak anti-HCV positive sera, 80 viraemic and anti-HCVpositive sera from patients infected with different subtypes and 10 sera from patients with HBV‐HCV or HIV‐HCV co-infections. Of 500 anti-HCV negative samples, 499 (99.8%) were negative with a cut-off index

We localized HCV C-100 protein in liver biopsies of 15 patients with chronic hepatitis C using immunohistochemistry. The results were compared to serum, tissue extract analysis of HCV RNA and in situ RT-PCR described in a previous study. HCV was detected in 80% of the sera tested, in 40% of the tissue extracts and in 80% and 60% of the tissue sections tested by immunohistochemistry and in situ RT-PCR respectively. Compared to the serum positive cases, 83% and 67% of the cases were respectively positive with immunohistochemistry and in situ RT-PCR and 41% were positive with tissue extract detection. Compared to the tissue extract positive cases, 25% and 50% of the cases were respectively positive with immunohistochemistry and in situ RT-PCR. Finally, 75% of the cases positive by immunohistochemistry were also positive by in situ RT-PCR. These results underline the complementarity of the different methods for the precise diagnosis of hepatitis C.

506 UNDETECTABLE HCV-RNA IN TELAPREVIR-TREATED PATIENTS: LOW CONCORDANCE BETWEEN TWO HIGHLY SENSITIVE REAL-TIME PCR ASSAYS J. Vermehren, A. Aghemo, K.-H. Peiffer, S. Susser, R. D’Ambrosio, D. Perner, L. Gerber, R. Soffredini, O. Weiland, S. Zeuzem, M. Colombo, C. Sarrazin. Medizinische Klinik 1, Klinikum der J. W. Goethe Universitaet, Frankfurt/Main, Germany; Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy; Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden E-mail: jvermehren@gmx.de

Using two sources of primary antibodies, we immunohisto-chemically stained hepatitis C virus-related antigen(s) on fixed-embedded liver specimens. These antigens were localized in the cytoplasm of hepato-cytes. The results obtained serologically correlated well with immunohisto-chemistry.

The complexity and cost of current diagnostics for hepatitis C virus (HCV) may act as a prevention to the scale-up of treatment in the developing world. Currently, ribonucleic acid (RNA)-polymerase chain reaction tests are the gold standard. However, there is potential for the use of simpler and cheaper antigen tests to confirm HCV infection in different clinical settings. We evaluated the sensitivity and specificity of antigen assays. This was compared with the reference-standard RNA assays. A subanalysis also assessed Architect core antigen test, which is the only commercially available antigen test on the market. In 24 datasets, evaluating HCV-antigen assays in 8136 samples, the percentage of HCV-antigen positive, HCV-RNA negative was 0.57%. The percentage HCV-antigen negative, HCV-RNA positive was 3.52%. There is strong evidence that antigen detection performs as well as RNA-based assays for HCV management. The use of antigen tests could improve access to HCV care in underresourced healthcare settings.