Epitope analysis of antibodies in Japanese to human cytomegalovirus phosphoprotein 150 with synthetic peptides.
Serological detection of antibodies specific to human cytomegalovirus (HCMV) is not reliable because the assay uses the whole HCMV protein fraction. Antigenic materials composed of well-characterized viral proteins are being tried for serodiagnosis in Europe. Epitopes of antibodies to HCMV phosphoprotein 150 (pp150) encoded by UL32 in Japanese individuals were investigated for comparison with the results in Europe. The major epitopes on amino acid residues 496 to 652 of HCMV pp150 were identified and the detection of antibodies with an enzyme-linked immunosorbent assay (ELISA) of synthetic peptides against the main epitopes was established. Fifteen seropositive and five seronegative serum samples for the epitope mapping and 131 seropositive and 50 seronegative samples for ELISA were investigated. Overlapping 15-mer peptides moving by two amino acids through V496-H652 were synthesized. The main epitope regions were V508-D530, L526-Q544, S536-D554, T616-G634, S624-P642, and L632-H652. When each peptide was conjugated with bovine serum albumin for ELISA, 80.9% of the seropositive samples were judged to be positive. The results of this study are the same as those for European sera, so the antigenic materials developed in Europe might be used to replace the whole HCMV protein fraction in Japanese.
Takahashi
Isao
I
Japanese Red Cross Aichi Blood Center, 539-3 Minami-yamaguchi, Seto, Aichi 489-8555, Japan. isaotaka@os.usen.ne.jp
Sugiura
Sayoko
S
Ohta
Hirotoshi
H
Ozawa
Kazuo
K
Kamiya
Tadashi
T
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000235:genetics / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology*; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000032:analysis* / Q000097:blood; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007564:Japan; D008969:Molecular Sequence Data; D010748:Phosphopeptides / Q000138:chemical synthesis / Q000235:genetics / Q000276:immunology*; D010750:Phosphoproteins; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*
D000596:Amino Acids; D000914:Antibodies, Viral; D000939:Epitopes; D010748:Phosphopeptides; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C056367:pp150 protein, Cytomegalovirus