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酶联免疫吸附试验中合成肽组合对改善犬内脏利什曼病和人内脏利什曼病的免疫诊断。

Improved canine and human visceral leishmaniasis immunodiagnosis using combinations of synthetic peptides in enzyme-linked immunosorbent assay.

机构信息

Departamento de Bioquímica e Imunologia, Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Belo Horizonte, Minas Gerais, Brasil.

出版信息

PLoS Negl Trop Dis. 2012;6(5):e1622. doi: 10.1371/journal.pntd.0001622. Epub 2012 May 22.

Abstract

BACKGROUND

Zoonotic visceral leishmaniasis (VL) is a severe infectious disease caused by protozoan parasites of the genus Leishmania and the domestic dogs are the main urban parasite reservoir hosts. In Brazil, indirect fluorescence antibody tests (IFAT) and indirect enzyme linked immunosorbent assay (ELISA) using promastigote extracts are widely used in epidemiological surveys. However, their sensitivity and specificity have often been compromised by the use of complex mixtures of antigens, which reduces their accuracy allowing the maintenance of infected animals that favors transmission to humans. In this context, the use of combinations of defined peptides appears favorable. Therefore, they were tested by combinations of five peptides derived from the previously described Leishmania diagnostic antigens A2, NH, LACK and K39.

METHODOLOGY/PRINCIPAL FINDINGS: Combinations of peptides derived A2, NH, LACK and K39 antigens were used in ELISA with sera from 44 human patients and 106 dogs. Improved sensitivities and specificities, close to 100%, were obtained for both sera of patients and dogs. Moreover, high sensitivity and specificity were observed even for canine sera presenting low IFAT anti-Leishmania antibody titers or from asymptomatic animals.

CONCLUSIONS/SIGNIFICANCE: The use of combinations of B cell predicted synthetic peptides derived from antigens A2, NH, LACK and K39 may provide an alternative for improved sensitivities and specificities for immunodiagnostic assays of VL.

摘要

背景

动物源内脏利什曼病(VL)是一种由原生动物利什曼原虫引起的严重传染病,而家犬是主要的城市寄生虫储存宿主。在巴西,间接荧光抗体试验(IFAT)和间接酶联免疫吸附试验(ELISA)使用前鞭毛体提取物广泛用于流行病学调查。然而,由于使用了复杂的抗原混合物,它们的敏感性和特异性经常受到影响,这降低了它们的准确性,从而允许感染动物的存在,这有利于向人类传播。在这种情况下,使用定义明确的肽组合似乎是有利的。因此,它们被测试了由五个肽的组合组成,这些肽衍生自先前描述的利什曼病诊断抗原 A2、NH、LACK 和 K39。

方法/主要发现:使用源自 A2、NH、LACK 和 K39 抗原的肽组合,对 44 名人类患者和 106 只狗的血清进行了 ELISA 检测。对患者和狗的血清都获得了接近 100%的更高敏感性和特异性。此外,即使对具有低 IFAT 抗利什曼抗体滴度或来自无症状动物的犬血清,也观察到了高敏感性和特异性。

结论/意义:使用源自 A2、NH、LACK 和 K39 抗原的 B 细胞预测合成肽的组合可能为 VL 的免疫诊断试验提供更高的敏感性和特异性的替代方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2cf5/3358334/f29569ab0d21/pntd.0001622.g001.jpg

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