Takahashi Isao, Sugiura Sayoko, Ohta Hirotoshi, Ozawa Kazuo, Kamiya Tadashi
Japanese Red Cross Aichi Blood Center, 539-3 Minami-yamaguchi, Seto, Aichi 489-8555, Japan.
Biosci Biotechnol Biochem. 2002 Nov;66(11):2402-5. doi: 10.1271/bbb.66.2402.
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.
人巨细胞病毒（HCMV）特异性抗体的血清学检测不可靠，因为该检测使用的是整个HCMV蛋白组分。欧洲正在尝试使用由特征明确的病毒蛋白组成的抗原材料进行血清学诊断。研究了日本个体中由UL32编码的HCMV磷蛋白150（pp150）抗体的表位，以便与欧洲的结果进行比较。确定了HCMV pp150氨基酸残基496至652上的主要表位，并建立了针对主要表位的合成肽酶联免疫吸附测定（ELISA）来检测抗体。研究了15份血清阳性和5份血清阴性的样本用于表位作图，以及131份血清阳性和50份血清阴性的样本用于ELISA。合成了在V496-H652区域以两个氨基酸移动的重叠15聚体肽。主要表位区域为V508-D530、L526-Q544、S536-D554、T616-G634、S624-P642和L632-H652。当将每种肽与牛血清白蛋白偶联用于ELISA时，80.9%的血清阳性样本被判定为阳性。本研究结果与欧洲血清的结果相同，因此欧洲开发的抗原材料可能可用于替代日本使用的整个HCMV蛋白组分。
