目的:为梅毒血清学诊断寻找新的血清诊断候选物。
方法:表达两种重组梅毒螺旋体蛋白Tp0100和Tp1016,纯化,并通过西方印迹鉴定。使用基于Tp0100的ELISA测试了总共600份临床血清样品,基于Tp1016的ELISA,和商业LICA梅毒TP试剂盒(ChIVD,北京,中国)。敏感性是通过测试来自临床诊断为原发性,次要,潜在的,和三级梅毒。通过从健康对照和潜在交叉反应性感染的个体中筛选260个样本来确定特异性。包括钩端螺旋体病,莱姆病,乙型肝炎,结核病,类风湿性关节炎,系统性红斑狼疮。Kappa(κ)值用于比较临床梅毒诊断与基于Tp0100的ELISA之间的一致性,基于Tp1016的ELISA,或LICA梅毒TP测试。
结果:以临床梅毒诊断为金标准,对于测试IgG抗体,Tp0100表现出95.6%的总体灵敏度和98.1%的特异性,而Tp1016仅表现出75.0%的总体灵敏度和79.6%的特异性。相比之下,LICA梅毒TP检测显示总体敏感性为97.6%,特异性为96.2%.此外,基于Tp0100的ELISA的总体一致性百分比和相应的κ值分别为96.7%(95%CI95.6%-97.8%)和0.93,基于Tp1016的ELISA为77.0%(95%CI74.3%-79.7%)和0.54,LICA梅毒TP试验为97.0%(95%CI96.0%-98.0%)和0.94,分别。
结论:重组梅毒螺旋体蛋白Tp0100有望作为梅毒血清学检测的新型诊断抗原。
OBJECTIVE: To discover novel serodiagnostic candidates for the serological diagnosis of syphilis.
METHODS: Two recombinant Treponema pallidum proteins Tp0100 and Tp1016 were expressed, purified, and identified by Western Blotting. A total of 600 clinical serum samples were tested with the Tp0100-based ELISA, the Tp1016-based ELISA, and the commercial LICA Syphilis TP kit (ChIVD, Beijing,
China). The sensitivities were determined by testing 340 samples from individuals with clinically diagnosed primary, secondary, latent, and tertiary syphilis. The specificities were determined by screening 260 samples from healthy controls and individuals with potentially cross-reactive infections, including leptospirosis, Lyme disease, hepatitis B, tuberculosis, rheumatoid arthritis, systemic lupus erythematosus. Kappa (κ) values were applied to compare the agreement between clinical syphilis diagnosis and the Tp0100-based ELISA, the Tp1016-based ELISA, or the LICA Syphilis TP test.
RESULTS: Using clinical syphilis diagnosis as the gold standard, Tp0100 exhibited an overall sensitivity of 95.6% and specificity of 98.1% for testing IgG antibody while Tp1016 demonstrated only an overall sensitivity of 75.0% and specificity of 79.6%. In contrast, the LICA Syphilis TP test revealed an overall sensitivity of 97.6% and specificity of 96.2%. In addition, the overall percent agreement and corresponding κ values were 96.7% (95% CI 95.6%-97.8%) and 0.93 for the Tp0100-based ELISA, 77.0% (95% CI 74.3%-79.7%) and 0.54 for the Tp1016-based ELISA, and 97.0% (95% CI 96.0%-98.0%) and 0.94 for the LICA Syphilis TP test, respectively.
CONCLUSIONS: The recombinant T. pallidum protein Tp0100 shows promise as a novel diagnostic antigen in the serological tests for syphilis.