Plachter B, Wieczorek L, Scholl B C, Ziegelmaier R, Jahn G
Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, Germany.
J Clin Microbiol. 1992 Jan;30(1):201-6. doi: 10.1128/jcm.30.1.201-206.1992.
Parts of the large phosphorylated tegument protein, pp150, of human cytomegalovirus (HCMV) were expressed in bacteria. The resulting fusion proteins were tested in a Western blot (immunoblot) assay for reactivity with a monoclonal antibody against pp150, with a polyspecific rabbit antiserum, and with human reconvalescent-phase sera. Those fusion proteins that performed well in the Western blot assay were used as antigens in enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against HCMV. Five different recombinant beta-galactosidase fusion proteins were evaluated by ELISA using 62 seropositive and 38 seronegative human serum samples. Of all the proteins tested, one peptide representing 162 amino acids of pp150 was superior to the others with regard to sensitivity and specificity. All sera known to be positive for antibodies against HCMV were identified by combining the results of the ELISAs with the different pp150 fusion proteins. Therefore, it appears that peptides from a single protein of HCMV might be sufficient to identify HCMV-seropositive individuals by recombinant ELISA.
人巨细胞病毒（HCMV）的大磷酸化被膜蛋白pp150的部分片段在细菌中表达。对产生的融合蛋白进行蛋白质印迹（免疫印迹）分析，检测其与抗pp150单克隆抗体、多特异性兔抗血清以及人恢复期血清的反应性。在蛋白质印迹分析中表现良好的那些融合蛋白被用作酶联免疫吸附测定（ELISA）中的抗原，用于检测抗HCMV抗体。使用62份血清阳性和38份血清阴性的人血清样本，通过ELISA评估了五种不同的重组β-半乳糖苷酶融合蛋白。在所有测试的蛋白中，一种代表pp150 162个氨基酸的肽段在敏感性和特异性方面优于其他肽段。通过将ELISA与不同的pp150融合蛋白的结果相结合，鉴定出所有已知抗HCMV抗体呈阳性的血清。因此，看来来自HCMV单一蛋白的肽段可能足以通过重组ELISA鉴定HCMV血清阳性个体。
