- Email: [email protected]
P.002 Low occurrence of occult hepatitis B virus infection (HBV) in patients with liver cirrhosis due to hepatitis C virus (HCV) with or without hepatocellular carcinoma (HCC)
Posters: 01a. Hepatitis B - diagnosis
$61
Posters [-~~
01a. Hepatitis B - d i a g n o s i s
[P•
Low occurrence of occult hepatitis B virus infection (HBV) in patients with liver cirrhosis due to hepatitis C virus (HCV) with or without hepatocellular carcinoma (HCC)
O. Guillaud 1, I.A. Chemin 1 *, S. Mrani 1, K. M6nouar 1, O. Galy 1, E Chevallier2, M. Trabeau 2, B. Seign~res 3, M. Les~n~chal 4, E Zoulim 1, C. Tr~po 1. 1U 271, INSERM, Lyon 69003; 2Laboratoire de virologie, H6pital de la Croix Rousse, Lyon 69317; 3Laboratoire, BioM6rieux, Marcy I'Etoile 69280; 4Laboratoire, BioM6rieux, Lyon 69317, France
R.S.S.M. Alencar 1, J.R.R. Pinho 1 *, M.M.S. Gomes 2, EM. Malta 2, R. Sitnik 3, I.M.V.G. Mello 4, E.S. Mello 5, T. Bacchella 1, M.C.C. Machado 1, V.A.E Alves 5, EJ. Carrilho 1. 1Departamento de Gastroenterologia, Faculdade de Medicina da Universidade de S#o Paulo; 2Servi~o de Virologia, Instituto Adolfo Lutz; 3Departamento de Patologia Clinica, Hospital Israelita Albert Einstein; 4Laborat6rio de Imunologia Viral, Instituto Butantan; 5Departamento de Patologia, Faculdade de Medicina da Universidade de S#o Paulo, S#o Paulo, Brazil
Background and Objectives: The pathogenic role of occult HBV infection remains unclear, and data concerning longitudinal follow-up of occult HBV carriers with cryptogenetic hepatitis is seldom referenced. Methods: In this study, 26 patients (15 men, 11 women, median age 48 years) suffering from cryptogenetic hepatitis, were selected according to the results of an initial HBV Nested PCR in S and/or X regions, and available follow-up sera. Clinical, biological and histological information was collected retrospectively. Follow-up patient sera were tested with three different immunoassays, of which 2 were designed to detect the most commonly described HBsAg mutants. In parallel, viral load quantification was performed using a Light Cycler PCR method and primers in the C region. Results: Initially, mean AST, ALT, GGT, AP values were respectively at 1.1N. 1.3N. 3.2N and 1.2N. During a mean 45-month follow-up period, these biological abnormalities persisted on GGT and ALT and were found in 80% of the samples tested. 22/26 patients had either mild or no fibrosis and 6/26 had cirrhosis, but none showed severe necroinflammatory lesions. When available, paired biopsy specimens showed no inflammation and stable fibrosis levels over 36 months. Initially, none of the 26 patients presented positive HBsAg serology in any of the 3 immunoassay tests used. For most of the virological follow-up of 11 patients, the three HBsAg tests remained negative despite quantifiable viremia. Only one of these patients presented an HBsAg-positive sample with all three tests due to a virological rebound with a viral load of up to 107 copies/ml. Previously, this patient remained below 104 copies/ml without detectable HBsAg. During the follow-up, viral loads remained below 103 copies/ml for 12 patients, between 103 and 106 copies/ml for 12 patients and over 106 for 2 patients. Conclusion: follow-up study of the 26 occult HBV patients follow-up showed (1) Persistent biological abnormalities on GGT and ALT. (2) Lower viremia than classic HBV chronic infection usually under 103 copies/ml but sometimes up to 106 or 107 copies/ml). Viral load variations of several logs were observed over time, even within one week, suggesting a highly dynamic viral population. (4) HBsAg remained undetectable although some follow-up samples reached viremia levels of 104 to 105 copies/ml.
Background and Objectives: Occult hepatitis B virus (HBV) infection has been found among patients with HCV and HIV infections, hepatitis non A - E and hepatocellular carcinoma (HCC). The aim of this study was to evaluate the presence of occult HBV infection in patients with liver cirrhosis related to hepatitis C (with or without HCC) that underwent liver transplantation in S~o Paulo, Brazil. Methods: Serum and liver tissue samples from 50 patients with HCV-related liver cirrhosis that underwent liver transplantation at the Hospital das Clfnicas, University of S~o Paulo School of Medicine, during the period of 1993 to 2004 were divided into two groups: Group 1 (33, cirrhosis) and Group 2 (17, cirrhosis with HCC). All patients were HBsAg negative, had previous serum samples frozen at -20°C and liver tissue explanted in paraffin, and did not present concomitant cholestatic, metabolic nor autoimmune liver diseases. The following variables were collected: gender, age, biochemical and coagulation laboratory tests and HBV serology (HBsAg, antiHBc total, anti-HBs). Among clinical data, ascites and encephalopathy were collected to determine Child and MELD indexes. Liver fragments were reviewed by specialized pathologist and classified using Ishak's Score and the Brazilian Society of Pathology and Hepatology Classification for chronic hepatitis and Edmondson and Steiner Classification for HCC. HBV DNA was detected in serum and in stored paraffin blocks of liver tissue (tumoral and non-tumoral) by the polymerase chain reaction (PCR) using in house (gene C) and real time PCR (gene S). External standard controls were run in parallel in all samples to check the quality of the extracted DNA. Results: All samples with or without tumoral liver tissue and serum were negative for HBV DNA using in house PCR. By real time PCR, only one case from Group 2 was HBVDNA positive in serum (male, 66 years old, isolated anti-HBc total positive and HCC). In the tumoral and non-tumoral liver tissues, there were two indeterminate HBV DNA cases among Group 2 patients as the standard control (beta actin) could not be amplified. All samples for Group 1 patients were negative for HBV DNA using both techniques Conclusion: This study showed an extremely low rate of occult hepatitis B virus infection among Brazilian HCV cirrhotic patients with or without HCC. Support: Alves de Queiroz Family Fund for Research, FAPESR
Characterization of occult HBV carriers according to a retrospective longitudinal follow-up of 26 patients
[-~~
Clinical relevance of the COBAS TaqMan HBV assay
M. Martinot-Peignoux 1 *, S. Maylin 2, J. Masurel 3, A. Rozenberg 3, Iq Marcellin 1. l lnserm U773 - CRB3; 2Dimpi INSERM U776-CRB3, H6pital Beaujon, Clichy, France, 3U-Paris Descartes, HSpital Cochin, Paris, France
Background and Objectives: Our aim was to assess clinical relevance of the introduction of the COBAS TaqMan HBV Assay in a routine laboratory Methods: 680 serum samples from patients with chronic hepatitis B undetectable serum HBV DNA with the VERSANT HBVDNA Assay v3.0 (bDNA) (Bayer Healthcare-Diagnostics; threshold 385 IU/ml) were routinely tested with (1) 536 samples with COBAS Amplicor HBV MONITOR assay, (Roche; threshold 501U/ml) and
$61
Posters [-~~
01a. Hepatitis B - d i a g n o s i s
[P•
Low occurrence of occult hepatitis B virus infection (HBV) in patients with liver cirrhosis due to hepatitis C virus (HCV) with or without hepatocellular carcinoma (HCC)
O. Guillaud 1, I.A. Chemin 1 *, S. Mrani 1, K. M6nouar 1, O. Galy 1, E Chevallier2, M. Trabeau 2, B. Seign~res 3, M. Les~n~chal 4, E Zoulim 1, C. Tr~po 1. 1U 271, INSERM, Lyon 69003; 2Laboratoire de virologie, H6pital de la Croix Rousse, Lyon 69317; 3Laboratoire, BioM6rieux, Marcy I'Etoile 69280; 4Laboratoire, BioM6rieux, Lyon 69317, France
R.S.S.M. Alencar 1, J.R.R. Pinho 1 *, M.M.S. Gomes 2, EM. Malta 2, R. Sitnik 3, I.M.V.G. Mello 4, E.S. Mello 5, T. Bacchella 1, M.C.C. Machado 1, V.A.E Alves 5, EJ. Carrilho 1. 1Departamento de Gastroenterologia, Faculdade de Medicina da Universidade de S#o Paulo; 2Servi~o de Virologia, Instituto Adolfo Lutz; 3Departamento de Patologia Clinica, Hospital Israelita Albert Einstein; 4Laborat6rio de Imunologia Viral, Instituto Butantan; 5Departamento de Patologia, Faculdade de Medicina da Universidade de S#o Paulo, S#o Paulo, Brazil
Background and Objectives: The pathogenic role of occult HBV infection remains unclear, and data concerning longitudinal follow-up of occult HBV carriers with cryptogenetic hepatitis is seldom referenced. Methods: In this study, 26 patients (15 men, 11 women, median age 48 years) suffering from cryptogenetic hepatitis, were selected according to the results of an initial HBV Nested PCR in S and/or X regions, and available follow-up sera. Clinical, biological and histological information was collected retrospectively. Follow-up patient sera were tested with three different immunoassays, of which 2 were designed to detect the most commonly described HBsAg mutants. In parallel, viral load quantification was performed using a Light Cycler PCR method and primers in the C region. Results: Initially, mean AST, ALT, GGT, AP values were respectively at 1.1N. 1.3N. 3.2N and 1.2N. During a mean 45-month follow-up period, these biological abnormalities persisted on GGT and ALT and were found in 80% of the samples tested. 22/26 patients had either mild or no fibrosis and 6/26 had cirrhosis, but none showed severe necroinflammatory lesions. When available, paired biopsy specimens showed no inflammation and stable fibrosis levels over 36 months. Initially, none of the 26 patients presented positive HBsAg serology in any of the 3 immunoassay tests used. For most of the virological follow-up of 11 patients, the three HBsAg tests remained negative despite quantifiable viremia. Only one of these patients presented an HBsAg-positive sample with all three tests due to a virological rebound with a viral load of up to 107 copies/ml. Previously, this patient remained below 104 copies/ml without detectable HBsAg. During the follow-up, viral loads remained below 103 copies/ml for 12 patients, between 103 and 106 copies/ml for 12 patients and over 106 for 2 patients. Conclusion: follow-up study of the 26 occult HBV patients follow-up showed (1) Persistent biological abnormalities on GGT and ALT. (2) Lower viremia than classic HBV chronic infection usually under 103 copies/ml but sometimes up to 106 or 107 copies/ml). Viral load variations of several logs were observed over time, even within one week, suggesting a highly dynamic viral population. (4) HBsAg remained undetectable although some follow-up samples reached viremia levels of 104 to 105 copies/ml.
Background and Objectives: Occult hepatitis B virus (HBV) infection has been found among patients with HCV and HIV infections, hepatitis non A - E and hepatocellular carcinoma (HCC). The aim of this study was to evaluate the presence of occult HBV infection in patients with liver cirrhosis related to hepatitis C (with or without HCC) that underwent liver transplantation in S~o Paulo, Brazil. Methods: Serum and liver tissue samples from 50 patients with HCV-related liver cirrhosis that underwent liver transplantation at the Hospital das Clfnicas, University of S~o Paulo School of Medicine, during the period of 1993 to 2004 were divided into two groups: Group 1 (33, cirrhosis) and Group 2 (17, cirrhosis with HCC). All patients were HBsAg negative, had previous serum samples frozen at -20°C and liver tissue explanted in paraffin, and did not present concomitant cholestatic, metabolic nor autoimmune liver diseases. The following variables were collected: gender, age, biochemical and coagulation laboratory tests and HBV serology (HBsAg, antiHBc total, anti-HBs). Among clinical data, ascites and encephalopathy were collected to determine Child and MELD indexes. Liver fragments were reviewed by specialized pathologist and classified using Ishak's Score and the Brazilian Society of Pathology and Hepatology Classification for chronic hepatitis and Edmondson and Steiner Classification for HCC. HBV DNA was detected in serum and in stored paraffin blocks of liver tissue (tumoral and non-tumoral) by the polymerase chain reaction (PCR) using in house (gene C) and real time PCR (gene S). External standard controls were run in parallel in all samples to check the quality of the extracted DNA. Results: All samples with or without tumoral liver tissue and serum were negative for HBV DNA using in house PCR. By real time PCR, only one case from Group 2 was HBVDNA positive in serum (male, 66 years old, isolated anti-HBc total positive and HCC). In the tumoral and non-tumoral liver tissues, there were two indeterminate HBV DNA cases among Group 2 patients as the standard control (beta actin) could not be amplified. All samples for Group 1 patients were negative for HBV DNA using both techniques Conclusion: This study showed an extremely low rate of occult hepatitis B virus infection among Brazilian HCV cirrhotic patients with or without HCC. Support: Alves de Queiroz Family Fund for Research, FAPESR
Characterization of occult HBV carriers according to a retrospective longitudinal follow-up of 26 patients
[-~~
Clinical relevance of the COBAS TaqMan HBV assay
M. Martinot-Peignoux 1 *, S. Maylin 2, J. Masurel 3, A. Rozenberg 3, Iq Marcellin 1. l lnserm U773 - CRB3; 2Dimpi INSERM U776-CRB3, H6pital Beaujon, Clichy, France, 3U-Paris Descartes, HSpital Cochin, Paris, France
Background and Objectives: Our aim was to assess clinical relevance of the introduction of the COBAS TaqMan HBV Assay in a routine laboratory Methods: 680 serum samples from patients with chronic hepatitis B undetectable serum HBV DNA with the VERSANT HBVDNA Assay v3.0 (bDNA) (Bayer Healthcare-Diagnostics; threshold 385 IU/ml) were routinely tested with (1) 536 samples with COBAS Amplicor HBV MONITOR assay, (Roche; threshold 501U/ml) and