Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo-Escola Paulista de Medicina, Disciplina de Imunologia, Rua Botucatu, 862, 4º andar, São Paulo, 04023-900, Brazil.
Mycopathologia. 2012 Jul;174(1):1-10. doi: 10.1007/s11046-011-9518-3. Epub 2012 Jan 17.
Paracoccidioidomycosis (PCM) is a systemic granulomatous disease, endemic in Latin America, caused by the thermal dimorphic fungus Paracoccidioides brasiliensis. Although some fungal antigens have already been characterized and used for serological diagnosis, cross-reactions have been frequently observed. Thus, the examination of fungal forms in clinical specimens or isolation of P. brasiliensis by culture is still the most frequent method for the diagnosis of this mycosis. In this study, a random peptide phage display library was used to select mimotopes of P. brasiliensis, which were employed as antigens in an indirect enzyme-linked immunosorbent assay. The protective monoclonal antibody against experimental PCM (anti-gp75) was used as molecular target to screen a phage display library. That approach led to a synthetic peptide named P2, which was synthesized and tested against PCM patients' sera to check whether it was recognized. There was significant recognition of P2 by sera of untreated PCM patients when compared with normal human sera. Sera from treated PCM group, patients with other mycosis or co-infected with HIV had much lower recognition of P2 than untreated patient group. The test showed a sensitivity of 100 and 94.59% of specificity in relation to human sera control. These data indicate a potential use of P2 as diagnostic tool in PCM. Its application for serological diagnosis of PCM may contribute to the development and standardization of simpler, faster and highly reproducible immunodiagnostic tests at low cost.
球孢子菌病(PCM)是一种全身性肉芽肿疾病,流行于拉丁美洲,由热双相真菌巴西副球孢子菌引起。虽然已经鉴定出一些真菌抗原并用于血清学诊断,但经常观察到交叉反应。因此,检查临床标本中的真菌形态或通过培养分离巴西副球孢子菌仍然是诊断这种真菌病最常见的方法。在这项研究中,使用随机肽噬菌体展示文库来选择巴西副球孢子菌的模拟表位,这些模拟表位被用作间接酶联免疫吸附试验的抗原。针对实验性 PCM 的保护性单克隆抗体(抗-gp75)被用作分子靶标来筛选噬菌体展示文库。这种方法导致了一种名为 P2 的合成肽的产生,该肽被合成并针对 PCM 患者的血清进行测试,以检查其是否被识别。与正常人血清相比,未经治疗的 PCM 患者的血清对 P2 的识别有显著差异。该测试对人血清对照的敏感性为 100%,特异性为 94.59%。这些数据表明 P2 可能作为 PCM 的诊断工具。它在 PCM 的血清学诊断中的应用可能有助于开发和标准化更简单、更快和高度可重复的免疫诊断测试,成本低廉。