de Sá Queiroz Júlio Henrique Ferreira, Dos Santos Barbosa Marcelo, Miranda Lais Gonçalves Ortolani, de Oliveira Natasha Rodrigues, Dellagostin Odir Antônio, Marchioro Silvana Beutinger, Simionatto Simone
Laboratório de Pesquisa em Ciências da Saúde, Universidade Federal da Grande Dourados - UFGD, Rodovia Dourados - Itahum, km 12, Cidade Universitária, Dourados, MS 79804970 Brazil.
Núcleo de Biotecnologia, Universidade Federal de Pelotas, UFPel, Pelotas, RS Brazil.
Indian J Microbiol. 2022 Sep;62(3):419-427. doi: 10.1007/s12088-022-01017-w. Epub 2022 Apr 19.
The incidence of syphilis has increased alarmingly over the years. Its diagnosis continues to be a challenge, leading to the search for new alternative and effective methods. The objective of this study was to select and evaluate three recombinant proteins for potential use in syphilis serodiagnosis. Bioinformatics analysis was performed with three antigens (Tp0684, Tp0750, and Tp0792) to assess their physical, antigenic, and structural characteristics. The antigens were chemically synthesized, recombinant plasmids were expressed in BL21 Star™ (DE3), and the recombinant proteins were purified by nickel affinity chromatography. The antigenicity of the recombinant proteins was evaluated by western blotting and enzyme-linked immunosorbent assay (ELISA), using the sera from patients with primary and latent syphilis. In silico analysis indicated the antigenic potential once the exposed B cell epitopes were detected in the evaluated proteins. Sera from patients with primary and latent syphilis specifically recognized rTp0684, rTp0750, and rTp0792 recombinant antigens. Moreover, the rTp0684-ELISA receiver operating characteristic (ROC) analysis showed an area under the ROC curve of 0.99, indicating high diagnostic efficacy with 97.62% specificity and 95% sensitivity. In conclusion, rTp0684 showed better potential as an antigen for the development of syphilis serodiagnosis. Thus, bioinformatic analysis can be an important tool to guide the selection of antigens for serological diagnosis.
The online version contains supplementary material available at 10.1007/s12088-022-01017-w.
多年来梅毒的发病率急剧上升。其诊断仍然是一项挑战,这促使人们寻找新的替代有效方法。本研究的目的是筛选和评估三种重组蛋白在梅毒血清学诊断中的潜在应用。对三种抗原(Tp0684、Tp0750和Tp0792)进行了生物信息学分析,以评估它们的物理、抗原和结构特征。这些抗原通过化学合成,重组质粒在BL21 Star™(DE3)中表达,重组蛋白通过镍亲和层析纯化。使用一期梅毒和潜伏梅毒患者的血清,通过蛋白质印迹法和酶联免疫吸附测定(ELISA)评估重组蛋白的抗原性。计算机分析表明,在所评估的蛋白质中一旦检测到暴露的B细胞表位,就具有抗原潜力。一期梅毒和潜伏梅毒患者的血清特异性识别rTp0684、rTp0750和rTp0792重组抗原。此外,rTp0684-ELISA的受试者工作特征(ROC)分析显示ROC曲线下面积为0.99,表明具有高诊断效能,特异性为97.62%,敏感性为95%。总之,rTp0684作为梅毒血清学诊断抗原显示出更好的潜力。因此,生物信息学分析可以成为指导血清学诊断抗原选择的重要工具。
在线版本包含可在10.1007/s12088-022-01017-w获取的补充材料。