Recombinant Treponema pallidum antigens in syphilis serology
Recombinant Treponema pallidum antigens in syphilis serology
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DOI：
10.1016/s0171-2985(97)80070-8
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发表时间：
1997-03-01
期刊：
影响因子：
2.8
通讯作者：
Morenz, J
中科院分区：
文献类型：
--
作者：
Gerber, A;Krell, S;Morenz, J
梅毒螺旋体是梅毒的病原体，其特征是表面暴露的外膜蛋白稀少，细胞质膜相关脂蛋白含量高。在感染的所有阶段，都可以检测到对脂蛋白的强烈抗体反应。为了给梅毒诊断提供抗原，克隆了高免疫原性脂蛋白TpN17、TpN29-35(Tpd)、TpN44.5(TmpA)、TpN47和TpN35(TmpC)以及膜蛋白TpN39(BMP)。将聚合酶链式反应扩增的DNA插入到E.colt表达载体中，可高效表达抗原。N-末端六组氨酸序列可用于金属螯合亲和层析对融合蛋白的高效纯化。重组抗原用酶联免疫吸附试验进行检测。TpN17、TpN47和TpN44.5抗原具有较高的抗体效价。与单一抗原相比，三种抗原联合检测进一步提高了诊断的敏感性。在18例梅毒患者中，有17例在所有阶段都发现了抗体，而42例正常人血清则为无反应。24例莱姆型疏螺旋体病患者血清中未检测到交叉反应。
Treponema pallidum, the etiological agent of syphilis, is characterized by a paucity of surface exposed outer membrane proteins and a high content of cytoplasma membrane associated lipoproteins. At all stages of infection intense antibody responses against lipoproteins are detectable. In order to provide antigens for syphilis diagnosis the highly immunogenic lipoproteins TpN17, TpN29-35 (TpD), TpN44.5 (TmpA), TpN47, and TpN35 (TmpC) and the membrane protein TpN39 (BMP) were cloned. Insertion of PCR amplified DNA into an E. colt expression vector resulted in high level expression of antigens. N-terminal hexahistidine sequence allowed efficient purification of fusion proteins by metal chelate affinity chromatography. The recombinant antigens were tested in enzyme-linked immunosorbent assays. TpN17, TpN47, and TpN44.5 antigens showed high antibody titers. Assays with the three antigens combined resulted in a further improvement of diagnostic sensitivity in comparison with single antigens. Antibodies were found in 17 of 18 patients in all stages of syphilis, whereas 42 normal human sera were nonreactive. No cross-reactivity was detected in 24 sera of patients with Lyme borreliosis.