Analysis of HBs antigen negative variant of hepatitis B virus: unique substitutions, Glu129 to Asp and Gly145 to Ala in the surface antigen gene.
We analyzed the surface gene (S gene) of a hepatitis B virus (HBV) isolate with mutations of envelope protein that rendered it undetectable by both a monoclonal hepatitis B surface antigen (HBsAg) enzyme-linked immunosorbent assay (ELISA) and polyclonal HBsAg radioimmunoassay (RIA). Sequencing of independently cloned products of HBV polymerase chain reaction revealed several point mutations within the S gene. Rare substitution was identified both at positions 129 (glutamine to asparagine) and at position 145 (glycine to alanine) in the 'a' determinant region, which is considered to be within a larger antigenic area known as the major hydrophilic region (MHR). A computer-assisted analysis of protein secondary structure could not find any significant difference between this mutant and wild-type HBsAg. However, the substitution of substitution glycine to alanine at position 129 introduce a putative glycosylation site (Asn-Gly-Thr), which may interfere with the antigenicity of HbsAg. Also, HBV variant with substitution at position 145 (Gly to Ala) has been recently reported to be antigenically altered and to show impaired recognition by polyclonal hepatitis B hyperimmune globulin in vitro. These genetic mutations in the S gene inside MHR may allow to escape detection by standard HBsAg assays.
Koyanagi
T
T
Department of Medicine and Bioregulatory Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Nakamuta
M
M
Sakai
H
H
Sugimoto
R
R
Enjoji
M
M
Koto
K
K
Iwamoto
H
H
Kumazawa
T
T
Mukaide
M
M
Nawata
H
H
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D005188:False Negative Reactions; D005814:Genes, Viral; D014644:Genetic Variation; D006031:Glycosylation; D006514:Hepatitis B Surface Antigens / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D019694:Hepatitis B, Chronic / Q000175:diagnosis / Q000821:virology; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D017354:Point Mutation; D017433:Protein Structure, Secondary
D000939:Epitopes; D006514:Hepatitis B Surface Antigens