Detection of hepatitis B virus core mutants by PCR-RFLP in chronically infected patients.
HBV chronic infection is an important health problem. The HBV core antigen carries several epitopes for T and B cell recognition and the immune response is crucial for determining the outcome of viral infection. Using PCR-RFLP several point mutations were detected in the HBV core ORF of HBV extracted from the serum of 140 chronically infected patients and 86 samples from another 37 patients followed-up in a longitudinal study. Mutations at position 2248 and 2147 (A3) and at 2038 (M2) were found most frequently. The wild type core genotype was found in about 50% of the samples. PCR-RFLP results were confirmed by direct sequencing of amplified products from HBV DNA present in chronically infected patients. The method is rapid and reliable and may be particularly useful for a rapid detection of viral mutants in a large number of patients.
Immunology Department, Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Campo de Santana 130, 1169-056 Lisbon, Portugal. imunologia@fcm.unl.pt
M F
MF
Barros
P A
PA
Rodrigues
S R
SR
Matias
L M
LM
Carreira
J A
JA
Machado-Caetano
D000293:Adolescent; D000328:Adult; D000368:Aged; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000097:blood; D005260:Female; D005838:Genotype; D006512:Hepatitis B Core Antigens / Q000097:blood / Q000737:chemistry / Q000235:genetics*; D006515:Hepatitis B virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D019694:Hepatitis B, Chronic / Q000821:virology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA
D004279:DNA, Viral; D006512:Hepatitis B Core Antigens