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Synthetic Peptides Mimic gp75 from Paracoccidioides brasiliensis in the Diagnosis of Paracoccidioidomycosis

机译:巴西副球菌模拟肽gp75在副球菌病诊断中的应用

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摘要

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.
机译:副球孢子菌病(PCM)是一种系统性肉芽肿病,是拉丁美洲的一种地方性肉芽肿病,是由巴西双歧热病菌引起的。尽管一些真菌抗原已被鉴定并用于血清学诊断,但经常观察到交叉反应。因此,检查临床标本中的真菌形式或通过培养分离巴西假单胞菌仍然是诊断这种真菌病的最常用方法。在这项研究中,使用随机肽噬菌体展示文库来选择巴西假单胞菌的模拟表位,将其用作间接酶联免疫吸附测定的抗原。针对实验性PCM的保护性单克隆抗体(抗gp75)被用作分子靶标,以筛选噬菌体展示文库。这种方法产生了一种名为P2的合成肽,该肽经合成并针对PCM患者的血清进行了测试,以检查其是否被识别。与正常人血清相比,未经治疗的PCM患者血清对P2有明显的认识。与未治疗的患者组相比,治疗的PCM组,其他真菌病或合并感染HIV的患者对P2的识别率低得多。该测试显示出相对于人类血清对照的特异性为100和94.59%。这些数据表明P2作为PCM中诊断工具的潜在用途。它在PCM血清学诊断中的应用可能有助于以较低的成本开发更简单,更快速且可重复性更高的免疫诊断测试并使之标准化。

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