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KMID : 0350519950480040979
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Journal of Catholic Medical College
1995 Volume.48 No. 4 p.979 ~ p.987
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Detection of Precore Mutant Hepatitis B Virus, A1896, by Polymerase in Korean Patients with Hepatitis B Virus Infection
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Abstract
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Hepatitis B e antigen(HBeAg) is derived form the translation product of the precore and core region. High levels of secreted HBeAg are found in highly viremic carriers. Elimination of HBeAg is usually accompanied by a flare-up of immune
pathogenesis.
However, not all patients lose serum HBV DNA and some of them continue to have elevated transaminase. Recently various mutations in the precore region have been detected in most of these HBeAg negative patients with high level of viremia. The
most
common form is a G to A mutation (mutant A1896) at position 1896 from the unique EcoR I site, which changes codon 28 from TGG to TAG, which is a translational stop codon. The pathogenetic roles of these precore mutants are controversial primarily
due to
observations of limited numbers of cases. As sequencing may be time-consuming, various approaches have been tried to screen the mutant A1896 by polymerase chain rcaction (PCR) products. The hybridization with oligoprobe and the allele specific
PCR
were
tried, but the procedures were also difficult and time-consuming. In a view of clinical importance of mutant A1896, the simple and rapid detection methods of mutant A1896 will required. Thus, we developed a method for detecting the mutant A1896
by
PCR
in combination with restriction fragment length polymorphism (RFLP). The PCR products of PCR in combination with restriction fragment length polymorphism (PFLP). The PCR products of wild type hepatitis B virus and mutant A1896 were digested with
Sty I
and Bsu36 I, respectively. Sera of 84 patients with hepatitis B virus infection (81: chronic, 3: acute) were examined for estimation of the clinical significance of mutant A1896.
@ES The results were as follows :
@EN 1. In all of three cases with acute hepatitis B, mutant A1896 were not detected. Out of 81 patients with chronic liver disease, 30 patients (37.0%) were infected with wild type virus only, 19 patients (21.7%) with wild type virus and mutant
A1896,
and 32 patients (39.5%) with mutant A1896 only.
2. Out of 26 HBeAg positive patients with chronic hepatitis B infection, 21 patients (80.8%) were infected with wild type virus only, 2 patients (7.7%) with wild type virus and mutant A1896 and 3 patients (11.5%) with mutant A1896 only. Out of
55
HBeAg
negative patients with chronic hepatitis B infection, 9 patients (16.4%) were infected with wild type virus only, 17 patients (30.9%) with wild type virus and mutant A1896, and 29 patients (52.7%) with mutant A1896 only.
3. All of 9 patients with HBeAg positive and inactive liver disease were infected with wild type virus only. In the 35 patients with HBeAg negative and active iver disease, the prevalence of mutant A1896 (88.6%) was higher than that of wild type
virus
only (11.4%).
4. On sequencing, all 9 cases which PCR products were digested by Sty I were G on nucleotide 1896 (wild type) and all 6 cases which PCR products were digested by Bsu36 I were A on nucleotide 1896 (A1896).
We suggested that PCR with modified primer in combination with two RFLP would be simple, easy and fast method to detect mutant A1896.
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KEYWORD
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