Toledo-Machado Christina Monerat, de Avila Ricardo Andrez Machado, NGuyen Christophe, Granier Claude, Bueno Lilian Lacerda, Carneiro Claudia Martins, Menezes-Souza Daniel, Carneiro Rubens Antonio, Chávez-Olórtegui Carlos, Fujiwara Ricardo Toshio
Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP 486, Belo Horizonte 31270-901, MG, Brazil.
Departamento Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, CP 486, Belo Horizonte 31270-901, MG, Brazil.
Biomed Res Int. 2015;2015:401509. doi: 10.1155/2015/401509. Epub 2015 Jan 29.
ELISA and RIFI are currently used for serodiagnosis of canine visceral leishmaniasis (CVL). The accuracy of these tests is controversial in endemic areas where canine infections by Trypanosoma cruzi may occur. We evaluated the usefulness of synthetic peptides that were selected through phage display technique in the serodiagnosis of CVL. Peptides were chosen based on their ability to bind to IgGs purified from infected dogs pooled sera. We selected three phage clones that reacted only with those IgGs. Peptides were synthesized, polymerized with glutaraldehyde, and used as antigens in ELISA assays. Each individual peptide or a mix of them was reactive with infected dogs serum. The assay was highly sensitive and specific when compared to soluble Leishmania antigen that showed cross-reactivity with anti-T. cruzi IgGs. Our results demonstrate that phage display technique is useful for selection of peptides that may represent valuable synthetic antigens for an improved serodiagnosis of CVL.
酶联免疫吸附测定(ELISA)和间接荧光免疫法(RIFI)目前用于犬内脏利什曼病(CVL)的血清学诊断。在可能发生克氏锥虫感染犬的流行地区,这些检测方法的准确性存在争议。我们评估了通过噬菌体展示技术筛选出的合成肽在CVL血清学诊断中的实用性。根据肽与从感染犬混合血清中纯化的免疫球蛋白(IgG)结合的能力来选择肽。我们挑选了三个仅与那些IgG发生反应的噬菌体克隆。合成肽,用戊二醛聚合,用作ELISA检测中的抗原。每种单独的肽或它们的混合物都能与感染犬的血清发生反应。与显示出与抗克氏锥虫IgG交叉反应的可溶性利什曼原虫抗原相比,该检测具有高度敏感性和特异性。我们的结果表明,噬菌体展示技术有助于筛选出可能代表有价值的合成抗原的肽,以改进CVL的血清学诊断。