A detection method for point mutation in the precore region of human hepatitis B virus (HBV)-DNA using mutation-site-specific assay.
In the natural progression of acute active hepatitis and chronic active hepatitis in human hepatitis B virus (HBV)-infected patients, inactive hepatitis develops by seroconversion, which can be explained by the disappearance of HBe antigen. However, it has been found that in some patients with hepatitis, alanine aminotransferase levels undergo fluctuation even though their serum is negative for HBe antigen. In these patients, HBV-DNA has been detected in the serum and the HBV-DNA so detected has been considered a cause of worsening liver function. Most HBV-DNA detected in these cases has a point mutation from G to A at the 83rd base in the precore region. As a result of this point mutation, HBV is unable to produce HBe antigen. We have devised a sensitive polymerase chain reaction (PCR) method, a mutation-site-specific assay, for the detection of point mutations at the 83rd base in the precore region using a specific mutation-trapped oligonucleotide primer for the mutant HBV genome.
Kinoshita
M
M
Diagnostic Research Institute, Otsuka Pharmaceutical Co., Ltd., Tokushima, Japan.
Seno
T
T
Fukui
T
T
Shin
S
S
Tsubota
A
A
Kumada
H
H
D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D004996:Ethidium; D006509:Hepatitis B / Q000821:virology*; D006513:Hepatitis B e Antigens / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D004279:DNA, Viral; D006513:Hepatitis B e Antigens; D004996:Ethidium