Alban Silvana Maria, de Moura Juliana Ferreira, Thomaz-Soccol Vanete, Bührer Sékula Samira, Alvarenga Larissa Magalhães, Mira Marcelo Távora, Olortegui Carlos Chávez, Minozzo João Carlos
Department of Biotechnology and Bioprocess Engineering, Federal University of Parana, Curitiba, Parana, Brazil.
Basic Pathology Department, Federal University of Parana, Curitiba, Parana, Brazil.
PLoS One. 2014 Aug 29;9(8):e106222. doi: 10.1371/journal.pone.0106222. eCollection 2014.
BACKGROUND: The diagnosis of leprosy is primarily based on clinical manifestations, and there is no widely available laboratory test for the early detection of this disease, which is caused by Mycobacterium leprae. In fact, early detection and treatment are the key elements to the successful control of leprosy. METHODOLOGY/PRINCIPAL FINDINGS: Peptide ligands for antibodies from leprosy patients were selected from phage-displayed peptide libraries. Three peptide sequences expressed by reactive phage clones were chemically synthesized. Serological assays that used synthetic peptides were evaluated using serum samples from leprosy patients, household contacts (HC) of leprosy patients, tuberculosis patients and endemic controls (EC). A pool of three peptides identified 73.9% (17/23) of multibacillary (MB) leprosy patients using an enzyme-linked immunosorbent assay (ELISA). These peptides also showed some seroreactivities to the HC and EC individuals. The peptides were not reactive to rabbit polyclonal antisera against the different environmental mycobacteria. The same peptides that were conjugated to the carrier protein bovine serum albumin (BSA) induced the production of antibodies in the mice. The anti-peptide antibodies that were used in the Western blotting analysis of M. leprae crude extracts revealed a single band of approximately 30 kDa in one-dimensional electrophoresis and four 30 kDa isoforms in the two-dimensional gel. The Western blotting data indicated that the three peptides are derived from the same bacterial protein. CONCLUSIONS/SIGNIFICANCE: These new antigens may be useful in the diagnosis of MB leprosy patients. Their potentials as diagnostic reagents must be more extensively evaluated in future studies using a large panel of positive and negative sera. Furthermore, other test approaches using peptides should be assessed to increase their sensitivity and specificity in detecting leprosy patients. We have revealed evidence in support of phage-displayed peptides as promising biotechnological tools for the design of leprosy diagnostic serological assays.
背景：麻风病的诊断主要基于临床表现，目前尚无广泛应用的实验室检测方法用于早期检测由麻风分枝杆菌引起的该疾病。事实上，早期检测和治疗是成功控制麻风病的关键因素。 方法/主要发现：从噬菌体展示肽库中筛选麻风病患者抗体的肽配体。化学合成了反应性噬菌体克隆表达的三个肽序列。使用来自麻风病患者、麻风病患者的家庭接触者（HC）、结核病患者和地方性对照（EC）的血清样本，对使用合成肽的血清学检测进行了评估。使用酶联免疫吸附测定（ELISA），一组三种肽可识别73.9%（17/23）的多菌型（MB）麻风病患者。这些肽对HC个体和EC个体也表现出一些血清反应性。这些肽对针对不同环境分枝杆菌的兔多克隆抗血清无反应。与载体蛋白牛血清白蛋白（BSA）偶联的相同肽可诱导小鼠产生抗体。用于麻风杆菌粗提物蛋白质印迹分析的抗肽抗体在一维电泳中显示出一条约30 kDa的条带，在二维凝胶中显示出四条30 kDa的同工型条带。蛋白质印迹数据表明这三种肽源自同一细菌蛋白。 结论/意义：这些新抗原可能有助于多菌型麻风病患者的诊断。在未来使用大量阳性和阴性血清的研究中，必须更广泛地评估它们作为诊断试剂的潜力。此外，应评估使用肽的其他检测方法，以提高其在检测麻风病患者中的敏感性和特异性。我们已揭示证据支持噬菌体展示肽作为设计麻风病诊断血清学检测的有前景的生物技术工具。
BMC Infect Dis. 2013-1-25
Am J Trop Med Hyg. 2013-3-11
Int J Lepr Other Mycobact Dis. 2002-3
Proteomes. 2021-1-29
J Immunol Res. 2017-3-7
BMC Infect Dis. 2013-1-25
Nat Genet. 2009-11-1
Rev Soc Bras Med Trop. 2008
Clin Vaccine Immunol. 2009-3
Clin Vaccine Immunol. 2008-10
Clin Vaccine Immunol. 2007-11
Microbes Infect. 2006-7
Zotero 插件
