Detection of hepatitis B virus precore TAG mutant by an amplification-created restriction site method

A new method for detecting the hepatitis B virus (HBV) precore 1896 G-->A mutation is described. This mutation prevents hepatitis B e antigen production by introducing a TAG stop codon and has been associated with severe chronic and fulminant hepatitis. The method is based on a polymerase chain reaction (PCR) that creates a restriction enzyme (Bsu36I) cleavage site if the mutation is present. After incubation of the PCR product with Bsu36I and a subsequent agarose gel electrophoresis, the presence of the TAG mutant is revealed by an altered position of the DNA band. The method was compared with direct sequencing on 36 serum samples and correctly identified all samples containing mutant HBV. The TAG mutant was present in 17 cases (as mixed wild type and mutant virus in 4). Twelve of 18 patients with advanced liver disease confirmed by biopsy carried mutant HBV. This method of detecting HBV precore 1896 G-->A should be useful for evaluation and follow-up of patients and for prevalence studies.