- Email: [email protected]
Apolipoprotein(a) inhibits hepatitis C virus entry
S82
Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
and is usually associated with virus inactivation [3]. Thought hepatitis B virus infection stages are serologically clearly defined the interpretation of results is not always easy in practice. E.g. anti-HBc indicates usually exposure to HBV, but found in daily practice profile such as: ‘anti-HBc only’ may mean false positive result. It is very important to give clinicians as accurate results as possible, so they can assess diseases stage and apply appropriate treatment. Aim of this study was to compare the results obtained with ADVIA Centaur XP (Siemens Healthcare Diagnostics, USA) chemiluminescence immunoassays, currently used to determine hepatitis B serological markers in our lab, with the results obtained with two other automated platforms: LIAISON® XL (DiaSorin, Saluggia, Italy) and Elecsys Cobas (Roche Diagnostics, Switzerland). Materials and methods: Serum samples (patient samples and quality specimen – UK, NEQAS) have been tasted for the qualitative or quantitative detection of 6 hepatitis B virus serological markers with assays from three chemiluminescent automated systems mentioned above. Results: The correlation between the tests performed on LIAISON® XL and ADVIA Centaur XP (patient samples) was: 100% for HBsAg, 96.7% for anti-HBs, 100% for anti-HBcIgM, 98.4% for anti-HBc-t, 100% for HBeAg and 95.3% for anti-HBe. The correlation between the tests performed on Elecsys and ADVIA Centaur XP (patient samples) was: 98.2% for HBsAg, 95.8% for anti-HBs, 97.7% for anti-HBcIgM, 94% for anti-HBc, 99% for HBeAg and 96.6% for anti-HBe. Weak positive ‘anti-HBc only’ results by the ADVIA Centaur anti-HBc assay were negative by both LIASON XL and Elecsys Anti-HBc assay. Results for all quality specimen were the same by all platforms. Conclusion: All three systems demonstrated assays suitable for routine use.
Methods: A total of 71 plasma samples of patients entering our laboratory with the request for HCV viral load and HCV genotyping were collected. Samples were analyzed in parallel with the COBAS AmpliPrep/COBAS TaqMan system and the DxN VERIS system. The HCV genotype was determined using the VERSANT HCV Genotype Assay Kit (LIPA) SIEMENS. Results: Of the71 samples tested 18 were positive and overall there was a good correlation between the two techniques (r = 0.963) for the parameter studied. The majority of the samples were determined to be HCV genotype 1, specifically 1a and 1b (n = 13), while three were un-typed. Other genotypes identified included HCV genotype 4a/4c/4d (n = 1) and 2a/2c (n = 1). Comparison of the VERIS HCV assay and Roche HCV assay results for the HCV genotype 1 sample showed that there was a good correlation between methods. This was observed for both genotype 1a or 1b. However for HCV genotype’s 2a/2c and 4a/4c/4d a difference was observed between both methods (difference of −0.95 log cp/mL for 2a/2c and −1.25 log cp/mL for 4a/4c/4d). Conclusion: The results indicate that the HCV viral load results generated on the DxN VERIS system were reproducible when compared to the reference method (COBAS). This together with the other features of the DxN VERIS system makes this device very useful in the daily routine diagnosis and monitoring of HCV viral load. The other features of the system include the ability to test samples from primary tubes and a bi-directional interface linking the DxN VERIS to the laboratory computer system (SIL). However, although it seems a coincidence, we should consider whether HCV genotypes 2 and 4 show discrepancies between the two techniques or is it a chance. A statistically significant number of samples of these genotypes should be tested to determine whether or not there is a discrepancy between the two methods. If this is confirmed then the cause should be determined.
Reference http://dx.doi.org/10.1016/j.jcv.2016.08.163 [1] D. Lavanchy, Hepatitis B virus epidemiology, disease burden, treatment, and current and emerging prevention and control measures, J. Viral Hepat. 11 (2) (2004) 97–107. [2] N. Hansen, G. Hay, S. Cowan, P. Jepsen, H. Bygum Krarup, N. Obel, et al., Hepatitis B prevalence in Denmark – an estimate based on nationwide registers and a national screening programme, as on 31 December 2007, Euro Surveill. Bull. Eur. Sur. Mal. Transm. Eur. Commun. Dis. Bull. 18 (47) (2013). [3] Diagnosís of hepatitis B virus infection, 2016. www.uptodate.com.
http://dx.doi.org/10.1016/j.jcv.2016.08.162 Abstract no: 352 Presentation at ESCV 2016: Poster 123 Comparative study of DxN VERIS molecular diagnostics system and the COBAS AmpliPrep/COBAS TaqMan platform for the determination of viral load in Hepatitis C virus infected patients J.M. Molina ∗ , J. Córdoba, M.D. Gomez, J.L. Hontangas Hospital La Fe, Valencia, Spain Introduction: The ability to measure the hepatitis C virus viral load in HCV infected patients has been available for several years on molecular diagnostics platforms from a number of manufacturer’s. Among these systems the COBAS AmpliPrep/COBAS TaqMan manufactured by Roche Diagnostics is widely used. The Beckman Coulter DxN VERIS system is a more recent addition to the list of platforms available for such assays. This aim of this study was to compare the performance of the DxN VERIS system with that of the Roche system for the accurate determination of HCV viral load in HCV infected patients.
Abstract no: 40 Presentation at ESCV 2016: Poster 124 Apolipoprotein(a) inhibits hepatitis C virus entry C. Oliveira 1,∗ , C. Fournier 1 , V. Descamps 1 , V. Morel 1 , C.A. Scipione 2 , M.L. Koschinsky 2 , A. Boullier 3 , P. Marcelo 4 , J.M. Domon 5 , E. Brochot 1 , G. Duverlie 1 , C. Francois 1 , S. Castelain 1 , F. Helle 1 1
EA4294, Laboratoire de Virologie, Centre Universitaire de Recherche en Santé, Centre Hospitalier Universitaire et Université de Picardie Jules Verne, 80054 Amiens, France 2 Department of Chemistry & Biochemistry, University of Windsor, Windsor, ON, Canada 3 INSERM U-1088, Université de Picardie Jules Verne, Centre Hospitalier Universitaire Sud, 80054 Amiens, France 4 Plateforme ICAP, Centre Universitaire de Recherche en Santé, Université de Picardie Jules Verne, 80054 Amiens, France 5 EA3900 BIOPI, Biologie des Plantes et Innovation, UFR des Sciences, Université de Picardie Jules Verne, 33 rue St Leu, 80039 Amiens, France In the last two decades, the development of different cell-based models has greatly contributed to improve the knowledge of HCV life cycle. However, it is still impossible to grow primary HCV isolates from each genotype in cell culture. This would open new
Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
perspectives to investigate viral determinants responsible for the different natural course and treatment outcome of hepatitis C as well as to develop a vaccine. In this study we hypothesized that this hindrance could be due to the presence of inhibitory factors in patient serum. Combining polyethylene glycol precipitation, iodixanol gradient and size exclusion chromatography, we obtained a purified fraction enriched in inhibitory factors from a pool of HCV seronegative serums. Mass spectrometry analysis of this fraction identified apolipoprotein(a) (apo(a)) as a potential inhibitor of the early step of HCV life cycle. Apo(a) consists of ten kringle IV-like domains (KIV), one kringle V-like domain (KV) and a protease-like domain that are homologous to plasminogen domains. Each of the ten KIV domains is present in a single copy with the exception of KIV type 2 (KIV 2), which is encoded in a variable number of tandemly repeated copies by the apo(a) gene, which gives rise to several apo(a) size isoforms in the human population. In addition, in human serum, apo(a) covalently links to the Apolipoprotein B component of a low density lipoprotein via a disulfide bridge to form a lipoprotein(a). The inhibitory effect of apo(a) on HCV entry was confirmed using a recombinant virus derived from the JFH1 strain and supernatant of cells transfected with plasmids expressing apo(a) as well as purified recombinant isoforms of apo(a). Our results also suggest that the larger the protein is, the better the inhibition is. We are currently testing several deletion mutants of apo(a) to identify critical domains for the inhibitory activity and to decipher the mechanism of inhibition. Altogether, our results identify apo(a) as an additional component of the lipid metabolism modulating HCV infection. http://dx.doi.org/10.1016/j.jcv.2016.08.164 Abstract no: 5 Presentation at ESCV 2016: Poster 125 Method comparison of VIDAS® ANTI-HBS TOTAL II with three equivalent assays in the 5–40 IU/L range critical for HBV vaccine status establishment G. Bouchard 1,∗ , V. Mossuz 2 , L. Levet 1 , Y. Ataman-Önal 1 , L. Demonchaux 1 , A. Dugua 1 , S. Jares Ferrier 1 , H. Convert 1 , J.M. Dugua 1 1 2
bioMérieux, Marcy l´étoile, France CEA, Grenoble, France
Vaccine-induced protection from hepatitis B virus (HBV) infection is correlated with the presence of antibodies against HBs antigen and individuals with a response ≥10 IU/L are considered protected. Each anti-HBs assay detects parts of the polyclonal antiHBs response, depending on its capture phase design. Criteria important for assay performance are equivalent recognition of HBV subtypes ad/ay and metrological agreement at 10 UI/L cut-off. This later is not easy to achieve and discrepancies between assays were repeatedly reported. Around 10 UI/L, discrepant results can translate into opposite clinical decisions regarding revaccination. We have compared 4 anti-HBsT assays in order to determine the one with the fewest discrepancies in the 0–40 IU/L range relevant for revaccination decision. 99 samples of routine HBV vaccine follow-up were collected from 3 laboratories in Grenoble area (CEA, CHU and Oriade Noviale). The recruitment criterion was a first anti-HBs result between 0 and 40 IU/L from one of the VIDAS, Architect or Cobas assays. In addition to these, all samples were also tested with the Biorad assay to generate 4 anti-HBsT results for each sample. For result interpreta-
S83
tion, <10 IU/L was considered negative and ≥10 IU/L was positive. Inter-assay qualitative agreements were defined as follows: total agreement is 4/4 positive or negative results, partial agreement is one discrepant result out of 4 and no agreement is 2 vs 2. 54/99 of anti-HBsT results were in total agreement and 35/99 were characterized by only one discrepant assay out of 4. The discrepancies were the following: 4 for VIDAS with 2 relative false positives and 2 relative false negatives, 7 for Architect with 2 relative false positives and 5 relative false negatives, 11 for Biorad, all relative false negatives, 13 for Cobas with 9 relative false positives and 2 relative false negatives. 10/99 of results were indeterminate and showed no agreement (2 vs 2). Among 4 assays, VIDAS anti-HBs Total II had the fewest discrepancies around the 10 IU/L cut-off owing to excellent analytical characteristics and enabled the most reliable decisions for HBV revaccination. http://dx.doi.org/10.1016/j.jcv.2016.08.165 Abstract no: 59 Presentation at ESCV 2016: Poster 126 Study of HCV seroprevalence in adult population in the Czech Republic R. Chlibek 1 , V. Stepanova 2,∗ , L. Pliskova 3 , J. Smetana 1 , S. Plisek 4 1
Department of Epidemiology, Faculty of Military Health Sciences, University of Defence, Hradec Kralove, Czech Republic 2 Inst. of Clin. Microbiology, University Hospital and Faculty of Medicine of Charles University, Hradec Kralove, Czech Republic 3 Inst. of Clin. Biochemistry and Diagnostics, University Hospital and Faculty of Medicine of Charles University, Hradec Kralove, Czech Republic 4 Clinic of Infectious Diseases, University Hospital and Faculty of Medicine of Charles University, Hradec Kralove, Czech Republic Background: Last official HCV seroprevalence survey in the Czech Republic (CZ) was performed by National Institute of Public Health (NIPH) in 2001 with HCV prevalence determined only 0.2%. Nevertheless chronic hepatitis C (VHC) is one of the frequent indication for liver transplantation in CZ (15.6%). According to the official report of NIPH up to 1000 of VHC cases and 82% VHC of all chronic hepatitis are reported annually in CZ (2015). The aim of our work was to determine the seroprevalence of HCV in CZ adult population, HCV viraemia and HCV genotype in HCV RNA positive persons and analyze the results as to the risk factors (i.v. drug users, health care workers) and estimate the number of persons with chronic hepatitis C in CZ. Materials and methods: The examined group included 3000 adult persons visiting in 02–09/2015 research centres of Hradec Kralove, Brno, Ceske Budejovice, males in 48.83%, females in 51.17%, age 18–90, median age 46 years. Anti-HCV antibodies were examined by 3rd generation test, CMIA (enzymatic immunoassay with chemiluminiscent detection), on Architect i2000, Abbott, with cut off S/CO <1 = negative (nonreactive); 1–2 = borderline reactive; >2 = positive (reactive). Samples with borderline reactivity were confirmed with immunoblot RIBA in NIPH. To determine viraemia all anti-HCV reactive samples were examined by RT–PCR, in HCV RNA positive samples genotypes were determined. Results: Of 3000 samples 50 were determined anti-HCV positive, seroprevalence of 1.67% (2.39% in males, 0.98% in females). 12 borderline reactive samples were confirmed negative by RIBA.
Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
and is usually associated with virus inactivation [3]. Thought hepatitis B virus infection stages are serologically clearly defined the interpretation of results is not always easy in practice. E.g. anti-HBc indicates usually exposure to HBV, but found in daily practice profile such as: ‘anti-HBc only’ may mean false positive result. It is very important to give clinicians as accurate results as possible, so they can assess diseases stage and apply appropriate treatment. Aim of this study was to compare the results obtained with ADVIA Centaur XP (Siemens Healthcare Diagnostics, USA) chemiluminescence immunoassays, currently used to determine hepatitis B serological markers in our lab, with the results obtained with two other automated platforms: LIAISON® XL (DiaSorin, Saluggia, Italy) and Elecsys Cobas (Roche Diagnostics, Switzerland). Materials and methods: Serum samples (patient samples and quality specimen – UK, NEQAS) have been tasted for the qualitative or quantitative detection of 6 hepatitis B virus serological markers with assays from three chemiluminescent automated systems mentioned above. Results: The correlation between the tests performed on LIAISON® XL and ADVIA Centaur XP (patient samples) was: 100% for HBsAg, 96.7% for anti-HBs, 100% for anti-HBcIgM, 98.4% for anti-HBc-t, 100% for HBeAg and 95.3% for anti-HBe. The correlation between the tests performed on Elecsys and ADVIA Centaur XP (patient samples) was: 98.2% for HBsAg, 95.8% for anti-HBs, 97.7% for anti-HBcIgM, 94% for anti-HBc, 99% for HBeAg and 96.6% for anti-HBe. Weak positive ‘anti-HBc only’ results by the ADVIA Centaur anti-HBc assay were negative by both LIASON XL and Elecsys Anti-HBc assay. Results for all quality specimen were the same by all platforms. Conclusion: All three systems demonstrated assays suitable for routine use.
Methods: A total of 71 plasma samples of patients entering our laboratory with the request for HCV viral load and HCV genotyping were collected. Samples were analyzed in parallel with the COBAS AmpliPrep/COBAS TaqMan system and the DxN VERIS system. The HCV genotype was determined using the VERSANT HCV Genotype Assay Kit (LIPA) SIEMENS. Results: Of the71 samples tested 18 were positive and overall there was a good correlation between the two techniques (r = 0.963) for the parameter studied. The majority of the samples were determined to be HCV genotype 1, specifically 1a and 1b (n = 13), while three were un-typed. Other genotypes identified included HCV genotype 4a/4c/4d (n = 1) and 2a/2c (n = 1). Comparison of the VERIS HCV assay and Roche HCV assay results for the HCV genotype 1 sample showed that there was a good correlation between methods. This was observed for both genotype 1a or 1b. However for HCV genotype’s 2a/2c and 4a/4c/4d a difference was observed between both methods (difference of −0.95 log cp/mL for 2a/2c and −1.25 log cp/mL for 4a/4c/4d). Conclusion: The results indicate that the HCV viral load results generated on the DxN VERIS system were reproducible when compared to the reference method (COBAS). This together with the other features of the DxN VERIS system makes this device very useful in the daily routine diagnosis and monitoring of HCV viral load. The other features of the system include the ability to test samples from primary tubes and a bi-directional interface linking the DxN VERIS to the laboratory computer system (SIL). However, although it seems a coincidence, we should consider whether HCV genotypes 2 and 4 show discrepancies between the two techniques or is it a chance. A statistically significant number of samples of these genotypes should be tested to determine whether or not there is a discrepancy between the two methods. If this is confirmed then the cause should be determined.
Reference http://dx.doi.org/10.1016/j.jcv.2016.08.163 [1] D. Lavanchy, Hepatitis B virus epidemiology, disease burden, treatment, and current and emerging prevention and control measures, J. Viral Hepat. 11 (2) (2004) 97–107. [2] N. Hansen, G. Hay, S. Cowan, P. Jepsen, H. Bygum Krarup, N. Obel, et al., Hepatitis B prevalence in Denmark – an estimate based on nationwide registers and a national screening programme, as on 31 December 2007, Euro Surveill. Bull. Eur. Sur. Mal. Transm. Eur. Commun. Dis. Bull. 18 (47) (2013). [3] Diagnosís of hepatitis B virus infection, 2016. www.uptodate.com.
http://dx.doi.org/10.1016/j.jcv.2016.08.162 Abstract no: 352 Presentation at ESCV 2016: Poster 123 Comparative study of DxN VERIS molecular diagnostics system and the COBAS AmpliPrep/COBAS TaqMan platform for the determination of viral load in Hepatitis C virus infected patients J.M. Molina ∗ , J. Córdoba, M.D. Gomez, J.L. Hontangas Hospital La Fe, Valencia, Spain Introduction: The ability to measure the hepatitis C virus viral load in HCV infected patients has been available for several years on molecular diagnostics platforms from a number of manufacturer’s. Among these systems the COBAS AmpliPrep/COBAS TaqMan manufactured by Roche Diagnostics is widely used. The Beckman Coulter DxN VERIS system is a more recent addition to the list of platforms available for such assays. This aim of this study was to compare the performance of the DxN VERIS system with that of the Roche system for the accurate determination of HCV viral load in HCV infected patients.
Abstract no: 40 Presentation at ESCV 2016: Poster 124 Apolipoprotein(a) inhibits hepatitis C virus entry C. Oliveira 1,∗ , C. Fournier 1 , V. Descamps 1 , V. Morel 1 , C.A. Scipione 2 , M.L. Koschinsky 2 , A. Boullier 3 , P. Marcelo 4 , J.M. Domon 5 , E. Brochot 1 , G. Duverlie 1 , C. Francois 1 , S. Castelain 1 , F. Helle 1 1
EA4294, Laboratoire de Virologie, Centre Universitaire de Recherche en Santé, Centre Hospitalier Universitaire et Université de Picardie Jules Verne, 80054 Amiens, France 2 Department of Chemistry & Biochemistry, University of Windsor, Windsor, ON, Canada 3 INSERM U-1088, Université de Picardie Jules Verne, Centre Hospitalier Universitaire Sud, 80054 Amiens, France 4 Plateforme ICAP, Centre Universitaire de Recherche en Santé, Université de Picardie Jules Verne, 80054 Amiens, France 5 EA3900 BIOPI, Biologie des Plantes et Innovation, UFR des Sciences, Université de Picardie Jules Verne, 33 rue St Leu, 80039 Amiens, France In the last two decades, the development of different cell-based models has greatly contributed to improve the knowledge of HCV life cycle. However, it is still impossible to grow primary HCV isolates from each genotype in cell culture. This would open new
Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
perspectives to investigate viral determinants responsible for the different natural course and treatment outcome of hepatitis C as well as to develop a vaccine. In this study we hypothesized that this hindrance could be due to the presence of inhibitory factors in patient serum. Combining polyethylene glycol precipitation, iodixanol gradient and size exclusion chromatography, we obtained a purified fraction enriched in inhibitory factors from a pool of HCV seronegative serums. Mass spectrometry analysis of this fraction identified apolipoprotein(a) (apo(a)) as a potential inhibitor of the early step of HCV life cycle. Apo(a) consists of ten kringle IV-like domains (KIV), one kringle V-like domain (KV) and a protease-like domain that are homologous to plasminogen domains. Each of the ten KIV domains is present in a single copy with the exception of KIV type 2 (KIV 2), which is encoded in a variable number of tandemly repeated copies by the apo(a) gene, which gives rise to several apo(a) size isoforms in the human population. In addition, in human serum, apo(a) covalently links to the Apolipoprotein B component of a low density lipoprotein via a disulfide bridge to form a lipoprotein(a). The inhibitory effect of apo(a) on HCV entry was confirmed using a recombinant virus derived from the JFH1 strain and supernatant of cells transfected with plasmids expressing apo(a) as well as purified recombinant isoforms of apo(a). Our results also suggest that the larger the protein is, the better the inhibition is. We are currently testing several deletion mutants of apo(a) to identify critical domains for the inhibitory activity and to decipher the mechanism of inhibition. Altogether, our results identify apo(a) as an additional component of the lipid metabolism modulating HCV infection. http://dx.doi.org/10.1016/j.jcv.2016.08.164 Abstract no: 5 Presentation at ESCV 2016: Poster 125 Method comparison of VIDAS® ANTI-HBS TOTAL II with three equivalent assays in the 5–40 IU/L range critical for HBV vaccine status establishment G. Bouchard 1,∗ , V. Mossuz 2 , L. Levet 1 , Y. Ataman-Önal 1 , L. Demonchaux 1 , A. Dugua 1 , S. Jares Ferrier 1 , H. Convert 1 , J.M. Dugua 1 1 2
bioMérieux, Marcy l´étoile, France CEA, Grenoble, France
Vaccine-induced protection from hepatitis B virus (HBV) infection is correlated with the presence of antibodies against HBs antigen and individuals with a response ≥10 IU/L are considered protected. Each anti-HBs assay detects parts of the polyclonal antiHBs response, depending on its capture phase design. Criteria important for assay performance are equivalent recognition of HBV subtypes ad/ay and metrological agreement at 10 UI/L cut-off. This later is not easy to achieve and discrepancies between assays were repeatedly reported. Around 10 UI/L, discrepant results can translate into opposite clinical decisions regarding revaccination. We have compared 4 anti-HBsT assays in order to determine the one with the fewest discrepancies in the 0–40 IU/L range relevant for revaccination decision. 99 samples of routine HBV vaccine follow-up were collected from 3 laboratories in Grenoble area (CEA, CHU and Oriade Noviale). The recruitment criterion was a first anti-HBs result between 0 and 40 IU/L from one of the VIDAS, Architect or Cobas assays. In addition to these, all samples were also tested with the Biorad assay to generate 4 anti-HBsT results for each sample. For result interpreta-
S83
tion, <10 IU/L was considered negative and ≥10 IU/L was positive. Inter-assay qualitative agreements were defined as follows: total agreement is 4/4 positive or negative results, partial agreement is one discrepant result out of 4 and no agreement is 2 vs 2. 54/99 of anti-HBsT results were in total agreement and 35/99 were characterized by only one discrepant assay out of 4. The discrepancies were the following: 4 for VIDAS with 2 relative false positives and 2 relative false negatives, 7 for Architect with 2 relative false positives and 5 relative false negatives, 11 for Biorad, all relative false negatives, 13 for Cobas with 9 relative false positives and 2 relative false negatives. 10/99 of results were indeterminate and showed no agreement (2 vs 2). Among 4 assays, VIDAS anti-HBs Total II had the fewest discrepancies around the 10 IU/L cut-off owing to excellent analytical characteristics and enabled the most reliable decisions for HBV revaccination. http://dx.doi.org/10.1016/j.jcv.2016.08.165 Abstract no: 59 Presentation at ESCV 2016: Poster 126 Study of HCV seroprevalence in adult population in the Czech Republic R. Chlibek 1 , V. Stepanova 2,∗ , L. Pliskova 3 , J. Smetana 1 , S. Plisek 4 1
Department of Epidemiology, Faculty of Military Health Sciences, University of Defence, Hradec Kralove, Czech Republic 2 Inst. of Clin. Microbiology, University Hospital and Faculty of Medicine of Charles University, Hradec Kralove, Czech Republic 3 Inst. of Clin. Biochemistry and Diagnostics, University Hospital and Faculty of Medicine of Charles University, Hradec Kralove, Czech Republic 4 Clinic of Infectious Diseases, University Hospital and Faculty of Medicine of Charles University, Hradec Kralove, Czech Republic Background: Last official HCV seroprevalence survey in the Czech Republic (CZ) was performed by National Institute of Public Health (NIPH) in 2001 with HCV prevalence determined only 0.2%. Nevertheless chronic hepatitis C (VHC) is one of the frequent indication for liver transplantation in CZ (15.6%). According to the official report of NIPH up to 1000 of VHC cases and 82% VHC of all chronic hepatitis are reported annually in CZ (2015). The aim of our work was to determine the seroprevalence of HCV in CZ adult population, HCV viraemia and HCV genotype in HCV RNA positive persons and analyze the results as to the risk factors (i.v. drug users, health care workers) and estimate the number of persons with chronic hepatitis C in CZ. Materials and methods: The examined group included 3000 adult persons visiting in 02–09/2015 research centres of Hradec Kralove, Brno, Ceske Budejovice, males in 48.83%, females in 51.17%, age 18–90, median age 46 years. Anti-HCV antibodies were examined by 3rd generation test, CMIA (enzymatic immunoassay with chemiluminiscent detection), on Architect i2000, Abbott, with cut off S/CO <1 = negative (nonreactive); 1–2 = borderline reactive; >2 = positive (reactive). Samples with borderline reactivity were confirmed with immunoblot RIBA in NIPH. To determine viraemia all anti-HCV reactive samples were examined by RT–PCR, in HCV RNA positive samples genotypes were determined. Results: Of 3000 samples 50 were determined anti-HCV positive, seroprevalence of 1.67% (2.39% in males, 0.98% in females). 12 borderline reactive samples were confirmed negative by RIBA.