Muniz Ana Paula Mendes, Tolesano-Pascoli Graziela, Vieira Raíssa Brauner Kamla, Polli Mayara Garcia, Rodrigues Vinícius da Silva, Gonzaga Henrique Tomaz, Mamede Carla Cristine Neves, Da Cunha Nathalie Costa, Szabó Matias Juan Pablo, Yokosawa Jonny
Laboratory of Microorganisms of Cerrado (Brazilian Savannah), Department of Microbiology, Instituto De Ciências Biomédicas (ICBIM), Universidade Federal de Uberlândia (UFU), Uberlândia, Brazil.
Laboratory of Ixodology, Faculdade de Medicina Veterinária, UFU, Uberlândia, Brazil.
Exp Appl Acarol. 2023 Feb;89(2):317-327. doi: 10.1007/s10493-023-00776-5. Epub 2023 Feb 16.
Rickettsia rickettsii is the etiological agent of Rocky Mountain spotted fever, which is an important tick-borne zoonosis and, in Brazil, it causes Brazilian spotted fever, which has high lethality rate. This study aimed to evaluate a synthetic peptide corresponding to a segment of the outer membrane protein A (OmpA) as an antigen in a serological test for the diagnosis of rickettsial infections. The amino acid sequence of the peptide was selected by predicting B cell epitopes using B Cell Epitope Prediction (Immune Epitope Database and Analysis Resource) and Epitopia and OmpA sequences of Rickettsia rickettsii strain 'Brazil' and Rickettsia parkeri strains 'Maculatum 20' and 'Portsmouth'. A peptide with amino acid sequence common to both Rickettsia species was synthesized and arbitrarily named OmpA-pLMC. To evaluate this peptide in enzyme-linked immunosorbent assay (ELISA), serum samples of capybara (Hydrochoerus hydrochaeris), horse (Equus caballus), and opossum (Didelphis albiventris) that had been previously tested by indirect immunofluorescence assay (IFA) for rickettsial infection were separated into IFA-positive and IFA-negative groups and used in the assay. There were no significant differences in ELISA optical density (OD) values between IFA-positive and IFA-negative groups with horse samples. The mean OD values were significantly higher in the IFA-positive capybara serum samples (IFA-pos vs. IFA-neg = 2.389 ± 0.761 vs. 1.760 ± 0.840). However, receiver operating characteristic (ROC) curve analysis did not show significant diagnostic parameters. On the other hand, 12 out of 14 (85.7%) opossum samples of the IFA-positive group showed reactivity in ELISA, and this was significantly higher than of the IFA-negative group (0.7196 ± 0.440 vs. 0.2318 ± 0.098, respectively; 85.7% sensitivity, 100% specificity). Therefore, our results show that OmpA-pLMC has a potential to be used in immunodiagnostic assays to detect spotted fever group rickettsial infections.
立氏立克次体是落基山斑疹热的病原体,落基山斑疹热是一种重要的蜱传人畜共患病,在巴西,它会引发致死率很高的巴西斑疹热。本研究旨在评估一种对应于外膜蛋白A(OmpA)一段序列的合成肽,作为用于诊断立克次体感染的血清学检测中的一种抗原。该肽的氨基酸序列是通过使用B细胞表位预测工具(免疫表位数据库和分析资源)以及Epitopia,对立氏立克次体“巴西”菌株、帕克立克次体“黄斑20”菌株和“朴茨茅斯”菌株的OmpA序列进行B细胞表位预测而选择出来的。合成了一种在两种立克次体物种中都具有共同氨基酸序列的肽,并随意命名为OmpA-pLMC。为了在酶联免疫吸附测定(ELISA)中评估该肽,将先前通过间接免疫荧光测定(IFA)检测立克次体感染的水豚(Hydrochoerus hydrochaeris)、马(Equus caballus)和负鼠(Didelphis albiventris)的血清样本分为IFA阳性组和IFA阴性组,并用于该测定。马的样本在IFA阳性组和IFA阴性组之间的ELISA光密度(OD)值没有显著差异。IFA阳性的水豚血清样本的平均OD值显著更高(IFA阳性组与IFA阴性组分别为2.389±0.761和1.760±0.840)。然而,受试者工作特征(ROC)曲线分析并未显示出显著的诊断参数。另一方面,IFA阳性组的14份负鼠样本中有12份(85.7%)在ELISA中显示出反应性,这显著高于IFA阴性组(分别为0.7196±0.440和0.2318±0.098;敏感性为85.7%,特异性为100%)。因此,我们的结果表明,OmpA-pLMC有潜力用于免疫诊断测定,以检测斑疹热群立克次体感染。