Detection of the hepatitis B virus major pre-core mutation by the amplification refractory mutation system technique.
HBeAg/anti-HBe seroconversion is associated, in some patients affected by type B chronic active hepatitis (CAH), with the occurrence of HBV pre-core mutants characterized by a common G-A change at codon 83. Since this mutation has important clinical correlations, we tried to develop a fast and reliable PCR test based on the amplification refractory mutation system (ARMS) technique, which has been successfully used to identify point mutations associated with genetic diseases. Following this approached, we analysed HBV particles isolated from 7 patients with anti-HBe CAH and previously characterized by DNA sequencing. Sera containing only wild type or mutant HBV DNA or a combination of both showed a discrete amplification product only in the presence of the specific wild type or mutant upstream primer. These results confirm the efficacy of the ARMS technique in detecting in a rapid and specific fashion the most common and clinically relevant HBV pre-core mutation.
Gramegna
M
M
Fondazione Centro Studi di Patologia Molecolare Applicata alla Clinica, Milano, Italy.
Lampertico
P
P
Lobbiani
A
A
Colucci
G
G
D001483:Base Sequence; D003062:Codon / Q000235:genetics; D006509:Hepatitis B / Q000382:microbiology*; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics*; D006521:Hepatitis, Chronic / Q000382:microbiology*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011498:Protein Precursors / Q000235:genetics; D014758:Viral Core Proteins / Q000235:genetics*
D003062:Codon; D006512:Hepatitis B Core Antigens; D011498:Protein Precursors; D014758:Viral Core Proteins