Semiquantitative assessment of pre-core stop-codon mutant and wildtype hepatitis B virus during the course of chronic hepatitis B using a new PCR-based assay

Archives of Virology
U Protzer-KnolleG Gerken

Abstract

In most patients with chronic hepatitis B positive for antibodies (anti-HBe) to HBe antigen (HBeAg), a pre-core mutant hepatitis B virus (HBV) with a point-mutation at nt. 1896 can be isolated. Clinical significance of the mutant virus in chronic hepatitis B is not proven yet, and screening of large numbers of sera during different clinical courses of numerous patients is necessary. We therefore aimed to develop a fast and reliable assay, that allows to discriminate wildtype from nt. 1896 G-->A mutant HBV and to determine the ratio of mutant and wildtype HBV in patients' sera. A mutation specific polymerase chain reaction (ms PCR) with new primers served to distinguish nt. 1896 G-->A mutant from wildtype HBV. This msPCR proved to be more sensitive and specific than similar assays described previously. When compared to a dilution series of a cloned HBV-DNA standard, the amount of wildtype and nt. 1896 G-->A mutant HBV could be determined semiquantitatively. 10(2) to 10(7) copies of each HBV-DNA (equivalent to 10(5) to 10(10) copies of HBV-DNA/ml patients' serum) could be amplified with steadily increasing signals. MsPCR proved to be specific as 10(7) copies did not give an amplification signal if they did not match the respectiv...Continue Reading

References

Feb 1, 1992·Gastroenterology·N V NaoumovH Will
May 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·M R BrunettoH Will
Jul 1, 1994·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·K HamasakiS Nagataki

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Citations

Oct 23, 2010·Expert Review of Molecular Diagnostics·Perumal Vivekanandan, Om Vir Singh
Sep 14, 2004·Revista do Hospital das Clínicas·Marcelo El Khouri, Vera Aparecida dos Santos
Feb 3, 2016·Dental Materials Journal·Takashi KomabayashiYohji Imai
Jul 3, 2004·Molecular Diagnosis : a Journal Devoted to the Understanding of Human Disease Through the Clinical Application of Molecular Biology·Perumal VivekanandanThenmozhi Subramaniam

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