Nunes D S, Gonzaga H T, Ribeiro V S, Cunha-Júnior J P, Costa-Cruz J M
Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia,Av. Pará 1720,38400-902, Uberlândia, MG,Brazil.
Parasitology. 2017 Apr;144(4):426-435. doi: 10.1017/S0031182016001839. Epub 2016 Oct 17.
There is an increasing interest in improving neurocysticercosis (NCC) diagnosis through the search of new and alternative antigenic sources, as those obtained from heterologous antigens. The aim of this study was to obtain potential biomarkers for NCC diagnosis after gel filtration chromatography [gel filtration fraction (GFF)] from the total saline extract (SE) from Taenia saginata metacestodes, followed by protein identification and application in immunodiagnostic. SE and GFF proteic profiles were characterized in gel electrophoresis, and diagnostic performance was verified by testing 160 serum samples through enzyme-linked immunosorbent assay and immunoblotting. Sensitivity (Se), specificity (Sp) and other diagnostic parameters were calculated. Polypeptides of interest in the diagnosis of human NCC present at GFF were analysed by mass spectrometry (MS) and B-cell epitopes were predicted. GFF had the best diagnostic parameters: Se 93·3%; Sp 93%; AUC 0·990; LR+ = 13·42 and LR- = 0·07, and proved to be useful reacting with serum samples in immunoblotting. Proteic profile ranged from 64 to 68 kDa and enolase and calcium binding protein calreticulin precursor were identified after MS. The enolase and calcium-binding protein calreticulin precursor showed 18 and 10 predicted B-cell epitopes, respectively. In conclusion we identified important markers in the GFF with high efficiency to diagnose NCC.
通过寻找新的和替代的抗原来源(如从异源抗原中获得的那些)来改善神经囊尾蚴病(NCC)诊断的兴趣与日俱增。本研究的目的是从牛带绦虫中绦期幼虫的总盐水提取物(SE)经凝胶过滤色谱法[凝胶过滤级分(GFF)]后获得用于NCC诊断的潜在生物标志物,随后进行蛋白质鉴定并应用于免疫诊断。在凝胶电泳中对SE和GFF的蛋白质谱进行了表征,并通过酶联免疫吸附测定和免疫印迹检测160份血清样本验证了诊断性能。计算了敏感性(Se)、特异性(Sp)和其他诊断参数。通过质谱(MS)分析了GFF中存在的对人类NCC诊断有意义的多肽,并预测了B细胞表位。GFF具有最佳诊断参数:Se 93.3%;Sp 93%;AUC 0.990;LR+ = 13.42和LR- = 0.07,并且在免疫印迹中与血清样本反应时被证明是有用的。蛋白质谱范围为64至68 kDa,质谱鉴定后确定烯醇化酶和钙结合蛋白钙网蛋白前体。烯醇化酶和钙结合蛋白钙网蛋白前体分别显示出18个和10个预测的B细胞表位。总之,我们在GFF中鉴定出了对诊断NCC高效的重要标志物。