Comparison of serum hepatitis B virus replication markers in patients with chronic hepatitis B: studies on HBeAg/Anti-HBe system, viral dna polymerase and HBV-DNA

Pinho, João Renato Rebello; Fonseca, Luís Edmundo Pinto da; Song, Yu; Miyamoto, Yuriko; Carrilho, Flair José; Granato, Celso Francisco Hernandes; Silva, Luiz Caetano da

doi:10.1590/S0036-46651989000500006

Abstracts

The detection of HBV-DNA in serum by molecular hybridization is the most sensitive and specific marker of replication and infectivity of hepatitis B virus and currently is proposed as a routine diagnostic technique in the follow-up of HBV - related diseases. Comparing different techniques already described, we found that direct spotting of serum samples on nitrocellulose membranes under vacuum filtration, followed by denaturing and neutralizing washes is more practical, simple, sensible and reproducible. DNA polymerase assay using phosphonoformic acid as specific viral inhibitor has shown 86.8% of concordance with HBV-DNA detection, and so, it is an useful alternative in the follow-up of hepatitis B chronic patients. We found 19.2% HBeAg positive samples with no other markers of viral replication and no anti-HBe positive sample had detectable HBV-DNA. Discordance between the 2 systems have been extensively described, and we confirm this for the first time in our country. Molecular biological techniques are essential to determine the replication status of chronic hepatitis B patients.

Hepatitis B; Molecular hybridization; DNA polymerase; HBV-DNA; HBV replication markers; Chronic hepatitis B

A detecção do genoma do HBV no soro por hibridização molecular é o mais sensível e específico marcador da replicação e infectividade do HBV, sendo sua utilização proposta como técnica rotineira no acompanhamento de doenças relacionadas a este vírus. Comparando diferentes técnicas descritas anteriormente, escolhemos a deposição direta das amostras séricas sobre a membrana de nitrocelulose sob filtração a vácuo, seguida de banhos desnaturantes e neutralisantes como mais prática e simples, com sensibilidade equivalente. O ensaio da DNA polimerase usando ácido fosfonofórmico como inibidor virai específico mostrou 86.8% de concordância com a detecção direta do DNA viral, sendo, portanto, uma alternativa viável no acompanhamento de pacientes com hepatite crônica B. Encontramos 19,2% das amostras AgHBe positivos sem outros marcadores de replicação viral. Por outro lado, nenhuma amostra anti-HBe positiva teve HBV-DNA detectável. Discordância entre estes dois sistemas foi extensamente descrita, e confirmamos pela primeira vez este fato em pacientes com hepatite crônica B em nosso país. Técnicas de biologia molecular são, portanto, fundamentais na determinação da replicação viral em cada paciente.

ORIGINAL ARTICLES

Comparison of serum hepatitis B virus replication markers in patients with chronic hepatitis B: studies on HBeAg/Anti-HBe system, viral dna polymerase and HBV-DNA

Comparação dos marcadores sorológicos da replicação do vírus da hepatite B: estudos sobre o sistema AgHBe/anti-HBe, DNA polimerase virai e HBV-DNA

João Renato Rebello Pinho^I; Luís Edmundo Pinto da Fonseca^II; Yu Song^I; Yuriko Miyamoto^I; Flair José Carrilho^II; Celso Francisco Hernandes Granato^III; Luiz Caetano da Silva^II

^IMolecular Biology Branch, Division of Virology, Instituto Adolfo Lutz, São Paulo, SP, Brazil

^IIClinical Hepatology Branch (LIM 07), Department of Gastroenterology and Hepatology Laboratory (LIM 47), Instituto de Medicina Tropical, Faculdade de Medicina da Universidade de São Paulo, São Paulo, SP, Brazil

^IIIHepatitis Branch, Division of Virology, Instituto Adolfo Lutz, São Paulo, SP, Brazil

^{Adress for correspondence} Address for correspondence: Dr. Luiz Caetano da Silva Instituto de Medicina Tropical Faculdade de Medicina da Universidade de São Paulo Av. Dr. Enéas de Carvalho Aguiar, 470 CEP 05403 São Paulo, SP, Brazil

SUMMARY

The detection of HBV-DNA in serum by molecular hybridization is the most sensitive and specific marker of replication and infectivity of hepatitis B virus and currently is proposed as a routine diagnostic technique in the follow-up of HBV - related diseases. Comparing different techniques already described, we found that direct spotting of serum samples on nitrocellulose membranes under vacuum filtration, followed by denaturing and neutralizing washes is more practical, simple, sensible and reproducible.

DNA polymerase assay using phosphonoformic acid as specific viral inhibitor has shown 86.8% of concordance with HBV-DNA detection, and so, it is an useful alternative in the follow-up of hepatitis B chronic patients.

We found 19.2% HBeAg positive samples with no other markers of viral replication and no anti-HBe positive sample had detectable HBV-DNA. Discordance between the 2 systems have been extensively described, and we confirm this for the first time in our country. Molecular biological techniques are essential to determine the replication status of chronic hepatitis B patients.

Key words: Hepatitis B; Molecular hybridization; DNA polymerase; HBV-DNA; HBV replication markers; Chronic hepatitis B.

RESUMO

A detecção do genoma do HBV no soro por hibridização molecular é o mais sensível e específico marcador da replicação e infectividade do HBV, sendo sua utilização proposta como técnica rotineira no acompanhamento de doenças relacionadas a este vírus. Comparando diferentes técnicas descritas anteriormente, escolhemos a deposição direta das amostras séricas sobre a membrana de nitrocelulose sob filtração a vácuo, seguida de banhos desnaturantes e neutralisantes como mais prática e simples, com sensibilidade equivalente.

O ensaio da DNA polimerase usando ácido fosfonofórmico como inibidor virai específico mostrou 86.8% de concordância com a detecção direta do DNA viral, sendo, portanto, uma alternativa viável no acompanhamento de pacientes com hepatite crônica B.

Encontramos 19,2% das amostras AgHBe positivos sem outros marcadores de replicação viral. Por outro lado, nenhuma amostra anti-HBe positiva teve HBV-DNA detectável. Discordância entre estes dois sistemas foi extensamente descrita, e confirmamos pela primeira vez este fato em pacientes com hepatite crônica B em nosso país. Técnicas de biologia molecular são, portanto, fundamentais na determinação da replicação viral em cada paciente.

Full text available only in PDF format.

Texto completo disponível apenas em PDF.

ACKNOWLEDGEMENTS

We are grateful to Doctors José Carlos da Costa Maia and Maria Helena Juliani for the alpha - 32 P - dATP, and to Doctors Francis Galibert and Christian Trepo for the plasmid containing the cloned HBV genome, to whom we are deeply indebted.

Recebido para publicação em 13/6/1989.

1. BERNINGER, M.; HAMMER, M.; HOYER, B. & GERIN, J. L. - An assay for detection of the DNA genome of hepatitis B virus in serum. J. med. Virol., 9: 57-66, 1982.
2. BONINO, F.; HOYER, B. & NELSON, J. - Hepatitis B virus DNA in the sera of HBs antigen carriers: a marker of active hepatitis B virus replication in the liver. Hepatology, 1: 386 391, 1981.
3. BRECHOT, C; HADCHOUEL, M.; SCOTTO, J.; DEGOS, F.; CHARNAY, P.; TREPO, C. & TIOLLAIS, P. - Detection of hepatitis B virus DNA in liver and serum: a direct appraisal of the chronic carrier status. Lancet, 2: 756-768, 1981.
4. BRECHOT, C; DEGOS, P.; LUGASSY, C; THIERS, V.' ZAFRANI, S.; FRANCO, D.; BENHAMOU, J. P.; WANDS, J.; ISSELBACHER, K.; TIOLLAIS, P. & BERTHELOT, P. - Hepatitis B virus DNA in patients with chronic liver disease and negative tests for hepatitis B surface antigen. New Engl. J. Med., 312: 270-276, 1985.
5. CARRILHO, F. J. & DA SILVA, L. C. - Epidemiologia. In: DA SILVA, L. C, ed. - Hepatites agudas e crônicas. São Paulo, Sarvier, 1986. p. 47-69.
6. CHAUSSADE, S.; THIERS, V.; BOBOC, B.; HOUSSET, C; BRECHOT, C. & DEGOS, F. - L'antigene HBe et l'ADN du virus B chez les malades d'hepatite chronique active. Gastroenterol, clin. biol., 10: 744-747, 1986.
7. CHU, C.-M; KARAYIANNIS, P.; FOWLER, M. J. F.; MONJARDINO, J.; LIAW, Y.-F. & THOMAS, H. C. - Natural history of chronic hepatitis B virus infection in Taiwan: studies of hepatitis B virus DNA in serum. Hepatology, 5: 431-434, 1985.
8. CRAXI, A.; WELLER, I. V. D. & BASSENDINE, M. F. - Relationship between HBV-specific DNA polymerase and HBe antigen/antibody system in chronic HBV infection: factors determining selection of patients and outcome of antiviral therapy. Gut, 24: 143-147, 1983.
9. FEINMAN, S. V.; BERRIS, B.; GUHA, A.; SOOKNANAN, R.; BRADDLEY, D. W.; BOND, W. W. & MAYNARD, J. E. - DNA: DNA hybridization method for the diagnosis of hepatitis B infection. J. virol. Meth., 8: 199-206, 1984.
10. FIGUS, A.; BLUM, H. E.; VYAS, G. N.; DE VIRGILIS, S.; CAO, A.; LIPPI, M.; LAI, E. & BALESTRIERI, A. - Hepatitis B viral sequences in non-A, non-3 or hepatitis B virus related chronic liver disease. Hepatology, 4: 364-368, 1984.
11. GENESCA, J.; JARDI, R.; PRAT, S.; BUTI, M.; STEBAN. R.; HERNANDEZ, J. M.; LOPEZ VIVANCOS, J.; PUIG DOMENICH, P. & GUARDIA, J. - Valor de la determinación sérica del DNA del virus de la hepatitis B como marcador de replicación virica. Med. clin. (Barcelona), 87: 665-668, 1986.
12. GERBER, M. A. & THUNG, S. N. - Biology of disease: molecular and cellular pathology of hepatitis. Lab. Invest., 52: 572-590, 1985.
13. GRANATO, C. F. H. - Avaliação do grau de replicação viral em indivíduos aguda ou cronicamente infectados pelo vírus da hepatite B: estudo sobre o antígeno HBe. São Paulo, 1988. (Dissertação de mestrado - Faculdade de Medicina da Universidade de São Paulo).
14. GUST, I. D.; BURRELL, C. J.; COULEPIS, A. G.; ROBIN SON, W. S. & ZUCKERMAN, A. J. - Taxonomic classification of human hepatitis B virus. Intervirology, 25: 14-29, 1986.
15. HARRISON, T. J.; BAL, V.; WHEELER, E. G.; MEACOCK, J. J.; HARRISON, J. F. & ZUCKERMAN, A. J. - Hepatitis B virus DNA and e antigen in serum from blood donors in the United Kingdom positive for hepatitis B surface antigen. Brit. med. J., 290: 663-664, 1985.
16. KAPLAN, P. M.; GREENMAN, R. L.; GERIN, J. L.; PURCELL, R. H. & ROBINSON, W. S. - DNA polymerase associated with human hepatitis B antigen. J. Virol., 12: 995-1005, 1973.
17. KARAYIANNIS, P.; FOWLER, M. J. F.; LOK, A. S. F.; GREENFIELD, C; MONJARDINO, J. & THOMAS, H. C. - Detection of serum HBV-DNA by molecular hybridization: correlation with HBeAg/anti-HBe status, racial origin, liver histology and hepatocellular carcinoma. J. Hepat., 1: 99-106, 1985.
18. LIEBERMANN, H. M.; LEBRECQUE, D. P.; KEW, M. C; HADZIYANNIS, S. J. & SHAFRITZ, D. A. - Hepatologic detection of hepatitis B virus DNA directly in human serum by a simplified molecular hybridization test: comparison to HBeAganti-HBe status in HBsAg carriers. Hepatology, 3: 285-291, 1983.
19. LIN, H. J.; WU, P.; LAI, C. L.; & CHAK, W. - Micromethod for phosphonoformic inhibition assay of hepatitis B viral DNA polymerase. Clin. Chem., 30: 549-552, 1984.
20. MANIATIS, T.; FRITSCH, E. F. & SAMBROOK, J. - Molecular cloning: a laboratory manual. Cold Spring Har bor, NY, Cold Spring Harbor Laboratory, 1982.
21. MEINKOTH, J. & WAHL, G. - Review: hybridization of nucleic acids immobilized on solid supports. Analyt. Biochem., 138: 267-284, 1984.
22. MORACE, G.; VON DER HELM, K. & DEINHARDT, F. - Detection of hepatitis B virus DNA in serum by a rapid filtration-hybridization assay. J. virol. Meth., 12: 235-242, 1985.
23. NORDENFELT, E. I. & KJELLEN, L. - Dane particles, DNA polymerase, and e antigen in twodistinct categories ot hepatitis B antigen carriers. Intervirology, 5: 225-232, 1975.
24. RIGBY, P. W. J.; DIECKMAN, M.; RHODES, C. & BERG, P. - Labelling deoxyribonucleic acid to high specific activity in vitro by nick-translation with DNA polymerase I. J. molec. Biol., 113: 237-251, 1976.
25. RINTJES, P. J. M.; VAN DITZHUIJSEN, J.; VAN LOON, A. M.; VAN HAELST, V. J. G. M.; BRONKHORST, F. B. & YAP, S. H. - Hepatitis B virus DNA detected in formalin fixed liver specimens and its relation to serologic markers and histopathologic features in chronic liver disease. Amer. J. Path., 120: 411-418, 1985.
26. SCOTTO, J.; HADCHOUEL, M.; HERY, C; YVART, J.; TIOLLAIS, P. & BRECHOT, C. - Detection of hepatitis B virus DNA in serum by a simple spot hybridization technique: comparison with results for other viral markers. Hepatology, 3: 279-284, 1984.
27. STANDRING, D. N. & RUTTER, W. J. - The molecular analysis of hepatitis B virus. In: POPPER, H. & SCHAFFNER, F., ed. - Progress in liver diseases. Orlando, Grune & Stratton, 1986. v. 8, p. 311-331.
28. VAN DITZHUIJSEN, T. J. M.; SELTEN, G. C. M.; VAN LOON, A. M.; WOLTERS, G.; MATTHYSSEN, L. & YAP, S. H. - Detection of Hepatitis B virus DNA in serum and relation with IgM class anti-HBc titers in hepatitis B virus infection. J. med. Virol., 15: 49-56, 1985.
29. WALTER, E.: BLUM, H. E.: OFFENSPERGER, W. B.; ZESCHNIGK, C; OFFENSPERGER, S. & GEROK, W. - Spot-blot hybridization assay for the detection of hepatitis B virus DNA in serum: factors determining its sensitivity and specificity. Hepatology, 7: 557-562,1987.
30. WELLER, I. V. D.; FOWLER, M. J. F.; MONJARDINO, J. & THOMAS, H. C. - The detection of HBV-DNA in serum by molecular hybridization: a more sensitive method for the detection of complete HBV particles. J. med. Virol., 9: 273-280, 1982.
31. ZELDIS, J. B.; BENPORATH, E.; ENAT, R.; KIRSCH, K. & WANDS, J. - Correlation of HBV-DNA and monoclonal reactivity to HBsAg in serum of patients with HBV infection. J. virol. Meth., 14: 153-166, 1986.

Address for correspondence:

Dr. Luiz Caetano da Silva

Instituto de Medicina Tropical

Faculdade de Medicina da Universidade de São Paulo

Av. Dr. Enéas de Carvalho Aguiar, 470

CEP 05403 São Paulo, SP, Brazil

Publication Dates

Publication in this collection
17 Nov 2010
Date of issue
Oct 1989

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. BERNINGER, M.; HAMMER, M.; HOYER, B. & GERIN, J. L. - An assay for detection of the DNA genome of hepatitis B virus in serum. J. med. Virol., 9: 57-66, 1982.

[2] 2. BONINO, F.; HOYER, B. & NELSON, J. - Hepatitis B virus DNA in the sera of HBs antigen carriers: a marker of active hepatitis B virus replication in the liver. Hepatology, 1: 386 391, 1981.

[3] 3. BRECHOT, C; HADCHOUEL, M.; SCOTTO, J.; DEGOS, F.; CHARNAY, P.; TREPO, C. & TIOLLAIS, P. - Detection of hepatitis B virus DNA in liver and serum: a direct appraisal of the chronic carrier status. Lancet, 2: 756-768, 1981.

[4] 4. BRECHOT, C; DEGOS, P.; LUGASSY, C; THIERS, V.' ZAFRANI, S.; FRANCO, D.; BENHAMOU, J. P.; WANDS, J.; ISSELBACHER, K.; TIOLLAIS, P. & BERTHELOT, P. - Hepatitis B virus DNA in patients with chronic liver disease and negative tests for hepatitis B surface antigen. New Engl. J. Med., 312: 270-276, 1985.

[5] 5. CARRILHO, F. J. & DA SILVA, L. C. - Epidemiologia. In: DA SILVA, L. C, ed. - Hepatites agudas e crônicas. São Paulo, Sarvier, 1986. p. 47-69.

[6] 6. CHAUSSADE, S.; THIERS, V.; BOBOC, B.; HOUSSET, C; BRECHOT, C. & DEGOS, F. - L'antigene HBe et l'ADN du virus B chez les malades d'hepatite chronique active. Gastroenterol, clin. biol., 10: 744-747, 1986.

[7] 7. CHU, C.-M; KARAYIANNIS, P.; FOWLER, M. J. F.; MONJARDINO, J.; LIAW, Y.-F. & THOMAS, H. C. - Natural history of chronic hepatitis B virus infection in Taiwan: studies of hepatitis B virus DNA in serum. Hepatology, 5: 431-434, 1985.

[8] 8. CRAXI, A.; WELLER, I. V. D. & BASSENDINE, M. F. - Relationship between HBV-specific DNA polymerase and HBe antigen/antibody system in chronic HBV infection: factors determining selection of patients and outcome of antiviral therapy. Gut, 24: 143-147, 1983.

[9] 9. FEINMAN, S. V.; BERRIS, B.; GUHA, A.; SOOKNANAN, R.; BRADDLEY, D. W.; BOND, W. W. & MAYNARD, J. E. - DNA: DNA hybridization method for the diagnosis of hepatitis B infection. J. virol. Meth., 8: 199-206, 1984.

[10] 10. FIGUS, A.; BLUM, H. E.; VYAS, G. N.; DE VIRGILIS, S.; CAO, A.; LIPPI, M.; LAI, E. & BALESTRIERI, A. - Hepatitis B viral sequences in non-A, non-3 or hepatitis B virus related chronic liver disease. Hepatology, 4: 364-368, 1984.

[11] 11. GENESCA, J.; JARDI, R.; PRAT, S.; BUTI, M.; STEBAN. R.; HERNANDEZ, J. M.; LOPEZ VIVANCOS, J.; PUIG DOMENICH, P. & GUARDIA, J. - Valor de la determinación sérica del DNA del virus de la hepatitis B como marcador de replicación virica. Med. clin. (Barcelona), 87: 665-668, 1986.

[12] 12. GERBER, M. A. & THUNG, S. N. - Biology of disease: molecular and cellular pathology of hepatitis. Lab. Invest., 52: 572-590, 1985.

[13] 13. GRANATO, C. F. H. - Avaliação do grau de replicação viral em indivíduos aguda ou cronicamente infectados pelo vírus da hepatite B: estudo sobre o antígeno HBe. São Paulo, 1988. (Dissertação de mestrado - Faculdade de Medicina da Universidade de São Paulo).

[14] 14. GUST, I. D.; BURRELL, C. J.; COULEPIS, A. G.; ROBIN SON, W. S. & ZUCKERMAN, A. J. - Taxonomic classification of human hepatitis B virus. Intervirology, 25: 14-29, 1986.

[15] 15. HARRISON, T. J.; BAL, V.; WHEELER, E. G.; MEACOCK, J. J.; HARRISON, J. F. & ZUCKERMAN, A. J. - Hepatitis B virus DNA and e antigen in serum from blood donors in the United Kingdom positive for hepatitis B surface antigen. Brit. med. J., 290: 663-664, 1985.

[16] 16. KAPLAN, P. M.; GREENMAN, R. L.; GERIN, J. L.; PURCELL, R. H. & ROBINSON, W. S. - DNA polymerase associated with human hepatitis B antigen. J. Virol., 12: 995-1005, 1973.

[17] 17. KARAYIANNIS, P.; FOWLER, M. J. F.; LOK, A. S. F.; GREENFIELD, C; MONJARDINO, J. & THOMAS, H. C. - Detection of serum HBV-DNA by molecular hybridization: correlation with HBeAg/anti-HBe status, racial origin, liver histology and hepatocellular carcinoma. J. Hepat., 1: 99-106, 1985.

[18] 18. LIEBERMANN, H. M.; LEBRECQUE, D. P.; KEW, M. C; HADZIYANNIS, S. J. & SHAFRITZ, D. A. - Hepatologic detection of hepatitis B virus DNA directly in human serum by a simplified molecular hybridization test: comparison to HBeAganti-HBe status in HBsAg carriers. Hepatology, 3: 285-291, 1983.

[19] 19. LIN, H. J.; WU, P.; LAI, C. L.; & CHAK, W. - Micromethod for phosphonoformic inhibition assay of hepatitis B viral DNA polymerase. Clin. Chem., 30: 549-552, 1984.

[20] 20. MANIATIS, T.; FRITSCH, E. F. & SAMBROOK, J. - Molecular cloning: a laboratory manual. Cold Spring Har bor, NY, Cold Spring Harbor Laboratory, 1982.

[21] 21. MEINKOTH, J. & WAHL, G. - Review: hybridization of nucleic acids immobilized on solid supports. Analyt. Biochem., 138: 267-284, 1984.

[22] 22. MORACE, G.; VON DER HELM, K. & DEINHARDT, F. - Detection of hepatitis B virus DNA in serum by a rapid filtration-hybridization assay. J. virol. Meth., 12: 235-242, 1985.

[23] 23. NORDENFELT, E. I. & KJELLEN, L. - Dane particles, DNA polymerase, and e antigen in twodistinct categories ot hepatitis B antigen carriers. Intervirology, 5: 225-232, 1975.

[24] 24. RIGBY, P. W. J.; DIECKMAN, M.; RHODES, C. & BERG, P. - Labelling deoxyribonucleic acid to high specific activity in vitro by nick-translation with DNA polymerase I. J. molec. Biol., 113: 237-251, 1976.

[25] 25. RINTJES, P. J. M.; VAN DITZHUIJSEN, J.; VAN LOON, A. M.; VAN HAELST, V. J. G. M.; BRONKHORST, F. B. & YAP, S. H. - Hepatitis B virus DNA detected in formalin fixed liver specimens and its relation to serologic markers and histopathologic features in chronic liver disease. Amer. J. Path., 120: 411-418, 1985.

[26] 26. SCOTTO, J.; HADCHOUEL, M.; HERY, C; YVART, J.; TIOLLAIS, P. & BRECHOT, C. - Detection of hepatitis B virus DNA in serum by a simple spot hybridization technique: comparison with results for other viral markers. Hepatology, 3: 279-284, 1984.

[27] 27. STANDRING, D. N. & RUTTER, W. J. - The molecular analysis of hepatitis B virus. In: POPPER, H. & SCHAFFNER, F., ed. - Progress in liver diseases. Orlando, Grune & Stratton, 1986. v. 8, p. 311-331.

[28] 28. VAN DITZHUIJSEN, T. J. M.; SELTEN, G. C. M.; VAN LOON, A. M.; WOLTERS, G.; MATTHYSSEN, L. & YAP, S. H. - Detection of Hepatitis B virus DNA in serum and relation with IgM class anti-HBc titers in hepatitis B virus infection. J. med. Virol., 15: 49-56, 1985.

[29] 29. WALTER, E.: BLUM, H. E.: OFFENSPERGER, W. B.; ZESCHNIGK, C; OFFENSPERGER, S. & GEROK, W. - Spot-blot hybridization assay for the detection of hepatitis B virus DNA in serum: factors determining its sensitivity and specificity. Hepatology, 7: 557-562,1987.

[30] 30. WELLER, I. V. D.; FOWLER, M. J. F.; MONJARDINO, J. & THOMAS, H. C. - The detection of HBV-DNA in serum by molecular hybridization: a more sensitive method for the detection of complete HBV particles. J. med. Virol., 9: 273-280, 1982.

[31] 31. ZELDIS, J. B.; BENPORATH, E.; ENAT, R.; KIRSCH, K. & WANDS, J. - Correlation of HBV-DNA and monoclonal reactivity to HBsAg in serum of patients with HBV infection. J. virol. Meth., 14: 153-166, 1986.

Brasil

Brasil

Comparison of serum hepatitis B virus replication markers in patients with chronic hepatitis B: studies on HBeAg/Anti-HBe system, viral dna polymerase and HBV-DNA

Comparação dos marcadores sorológicos da replicação do vírus da hepatite B: estudos sobre o sistema AgHBe/anti-HBe, DNA polimerase virai e HBV-DNA

Abstracts

Publication Dates