Lillehoj E P, Alexander S S, Dubrule C J, Wiktor S, Adams R, Tai C C, Manns A, Blattner W A
Cambridge Biotech Corp., Rockville, Maryland 20850.
J Clin Microbiol. 1990 Dec;28(12):2653-8. doi: 10.1128/jcm.28.12.2653-2658.1990.
A recombinant protein derived from the gp21 region of the human T-cell leukemia virus type I (HTLV-I) env gene was synthesized in Escherichia coli and purified by reversed-phase high-performance liquid chromatography. The purified protein was free of contaminating bacterial proteins and retained reactivity with human HTLV-I- and HTLV-II-positive sera and a gp21 monoclonal antibody. An immunoblot procedure using the recombinant polypeptide in conjunction with native viral proteins was more sensitive than the conventional immunoblot and radioimmunoprecipitation confirmatory assays for detection of antibodies to HTLV-I and HTLV-II env-encoded gene products. The recombinant protein was equally reactive with sera from polymerase chain reaction-confirmed HTLV-I or HTLV-II infections. Furthermore, on the basis of the differential reactivities of gp21-positive sera with the HTLV-I p19 and p24 gag-encoded proteins, an algorithm was proposed to distinguish exposure to HTLV-I from exposure to HTLV-II. These results establish the utility of a modified immunoblot assay incorporating a recombinant envelope polypeptide as an alternative to existing HTLV-I-confirmatory assays.
一种源自人I型嗜T细胞白血病病毒（HTLV-I）env基因gp21区域的重组蛋白在大肠杆菌中合成，并通过反相高效液相色谱法进行纯化。纯化后的蛋白不含污染性细菌蛋白，且与人HTLV-I和HTLV-II阳性血清以及一种gp21单克隆抗体保持反应活性。使用重组多肽结合天然病毒蛋白的免疫印迹程序，在检测针对HTLV-I和HTLV-II env编码基因产物的抗体方面，比传统免疫印迹和放射免疫沉淀确证试验更灵敏。该重组蛋白与经聚合酶链反应确证的HTLV-I或HTLV-II感染患者的血清具有同等反应活性。此外，基于gp21阳性血清与HTLV-I p19和p24 gag编码蛋白的不同反应性，提出了一种算法来区分HTLV-I暴露与HTLV-II暴露。这些结果证实了一种改良免疫印迹试验的实用性，该试验采用重组包膜多肽作为现有HTLV-I确证试验的替代方法。
