Lenz A, von Hintzenstern J, Erlwein O, Ellinger S, Bröker M, Fleckenstein B, Jahn G
Institut für Klinische und Molekulare Virologie der Universität Erlangen-Nürnberg.
Klin Wochenschr. 1987 Nov 2;65(21):1042-7. doi: 10.1007/BF01726323.
Serologic testing for human immunodeficiency virus type 1 (HIV-1) is currently based on enzyme linked immunosorbent assay (ELISA) as screening method. Positive ELISA-results have to be confirmed by at least one second procedure such as Western blotting or immunofluorescence. To obtain new diagnostic reagents for confirmatory testing, we expressed viral antigens in procaryotic systems. Peptides representing epitopes of structural core (gag)- and envelope (env)-proteins of HIV were produced in E. coli as stable immunogenic beta-galactosidase fusion proteins. Recombinant proteins were taken for immunoblot-assays. The results of Western blotting with those fusion proteins were in general comparable with conventional ELISA, immunofluorescence, immunoblot with cell-culture derived virus and commercially available ELISA tests based on recombinant proteins. Immunoblots using recombinant transmembrane protein (gp41) derived polypeptide were more sensitive than the conventional procedure with purified virion proteins. Western blotting with recombinant fusionproteins provide reliable and inexpensive serodiagnostics without handling of infectious cell cultures.
目前，1型人类免疫缺陷病毒（HIV-1）的血清学检测是以酶联免疫吸附测定（ELISA）作为筛查方法。ELISA检测呈阳性的结果必须通过至少一种其他方法进行确认，如蛋白质印迹法或免疫荧光法。为了获得用于确证检测的新型诊断试剂，我们在原核系统中表达病毒抗原。代表HIV结构核心（gag）蛋白和包膜（env）蛋白表位的肽段在大肠杆菌中作为稳定的免疫原性β-半乳糖苷酶融合蛋白产生。重组蛋白用于免疫印迹分析。这些融合蛋白的蛋白质印迹结果总体上与传统ELISA、免疫荧光、细胞培养衍生病毒的免疫印迹以及基于重组蛋白的市售ELISA检测相当。使用重组跨膜蛋白（gp41）衍生多肽的免疫印迹比使用纯化病毒粒子蛋白的传统方法更灵敏。用重组融合蛋白进行蛋白质印迹提供了可靠且廉价的血清学诊断方法，无需处理具有传染性的细胞培养物。
