Jiang Chuanhao, Zhao Feijun, Xiao Jinhong, Zeng Tiebing, Yu Jian, Ma Xiaohua, Wu Haiying, Wu Yimou
Pathogenic Biology Institute, Medical College, University of South China, Hengyang, China.
Clin Vaccine Immunol. 2013 Oct;20(10):1563-8. doi: 10.1128/CVI.00122-13. Epub 2013 Aug 14.
Syphilis is a chronic infection caused by Treponema pallidum subsp. pallidum, and diagnosis with sensitive and specific methods is a challenging process that is important for its prevention and treatment. In the present study, we established a recombinant protein TpF1-based indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and a Western blot assay for human and rabbit sera. The 20-kDa recombinant protein TpF1 was detected by Western blotting performed with sera from rabbits immunized with recombinant TpF1 and infected with the T. pallidum Nichols strain and T. pallidum clinical isolates but was not detected by Western blotting with sera from uninfected rabbits. The sensitivity of the recombinant protein was determined by screening sera from individuals with primary, secondary, latent, and congenital syphilis (n = 82). The specificity of the recombinant protein was determined by screening sera from uninfected controls (n = 30) and individuals with potentially cross-reactive infections, including Lyme disease (n = 30) and leptospirosis (n = 5). The sensitivities of TpF1-based ELISAs were 93.3%, 100%, 100%, and 100% for primary, secondary, latent, and congenital syphilis, respectively, and the specificities were all 100% for sera from uninfected controls and individuals with potentially cross-reactive infections. In Western blot assays, the sensitivities and specificities of TpF1 for human sera were all 100%. The reactivities of TpF1 with syphilitic sera were proportional to the titers of the T. pallidum particle agglutination (TPPA) assay. These data indicate that the recombinant protein TpF1 is a highly immunogenic protein in human and rabbit infections and a promising marker for the screening of syphilis.
梅毒是由梅毒螺旋体苍白亚种引起的一种慢性感染,采用敏感且特异的方法进行诊断是一个具有挑战性的过程,对其预防和治疗至关重要重要。在本研究中,我们建立了基于重组蛋白TpF1的间接免疫球蛋白G(IgG)酶联免疫吸附测定(ELISA)以及用于检测人和兔血清的免疫印迹法。用重组TpF1免疫并感染梅毒螺旋体Nichols菌株和梅毒螺旋体临床分离株的兔血清进行免疫印迹检测,可检测到20 kDa的重组蛋白TpF1,但未感染的兔血清进行免疫印迹检测未检测到。通过筛查一期、二期、潜伏和先天性梅毒患者(n = 82)的血清来确定重组蛋白的敏感性。通过筛查未感染对照(n = 30)以及患有可能存在交叉反应感染(包括莱姆病,n = 30;钩端螺旋体病,n = 5)的个体的血清来确定重组蛋白的特异性。基于TpF1的ELISA对一期、二期、潜伏和先天性梅毒的敏感性分别为93.3%、100%、100%和100%,对未感染对照和可能存在交叉反应感染个体的血清特异性均为100%。在免疫印迹分析中,TpF1对人血清的敏感性和特异性均为100%。TpF1与梅毒血清的反应性与梅毒螺旋体颗粒凝集试验(TPPA)的滴度成正比。这些数据表明,重组蛋白TpF1在人和兔感染中是一种高度免疫原性的蛋白,是梅毒筛查的一个有前景的标志物。