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利什曼原虫分泌的铁超氧化物歧化酶的纯化及其在犬利什曼病诊断中的应用。

Leishmania infantum secreted iron superoxide dismutase purification and its application to the diagnosis of canine Leishmaniasis.

机构信息

Departamento de Parasitología, Facultad de Ciencias, Universidad de Granada, Severo Ochoa s/n, 18071 Granada, Spain.

出版信息

Comp Immunol Microbiol Infect Dis. 2013 Sep;36(5):499-506. doi: 10.1016/j.cimid.2013.05.004. Epub 2013 Jun 12.

Abstract

Leishmania spp. are digenetic parasites whose infection occurs inside the mononuclear phagocitary system. The iron superoxide dismutase secreted (Fe-SODe) by promastigotes of Leishmania spp. seems to plays an important role in the defense to environmental detoxification and neutralization of oxidative stress damage caused by reactive oxygen species (ROS) produced by macrophages during the infection. Parasites Fe-SODe is involved in establishing the infection and manifestation of Leishmaniasis. Its high immunogenicity makes it a useful molecular marker in diagnosing trypanosomatids infections. The aim of this study is demonstrate that purified Fe-SODe from Leishmania infantum is much more sensitive than un-purified Fe-SODe for diagnosis canine Leishmaniasis. We have purified a Fe-SODe of L. infantum using an ion exchange and a molecular sieve chromatographies and its application in diagnosis of canine Leishmaniasis was tested. One hundred and forty-five dogs' sera from Andalusia Autonomous Community, Spain were tested by ELISA and Western blot and the antigen Fe-SODe purified is compared with two different antigens: the total parasites soluble lysate and the unpurified Fe-SODe. To validate the results obtained using the Fe-SODe purified we tasted 10 L. infantum infected dogs' sera from Lombardy, Italy as positive control.

摘要

利什曼原虫属是双细胞寄生虫,其感染发生在单核吞噬细胞系统内。利什曼原虫属的前鞭毛体分泌的铁超氧化物歧化酶(Fe-SODe)似乎在防御环境解毒和中和由感染期间巨噬细胞产生的活性氧(ROS)引起的氧化应激损伤方面发挥重要作用。寄生虫的 Fe-SODe 参与了利什曼病的感染和表现。其高度的免疫原性使其成为诊断锥虫感染的有用分子标志物。本研究旨在证明从婴儿利什曼原虫中纯化的 Fe-SODe 比未纯化的 Fe-SODe 更敏感,可用于诊断犬利什曼病。我们使用离子交换和分子筛层析法从婴儿利什曼原虫中纯化了 Fe-SODe,并测试了其在诊断犬利什曼病中的应用。使用 ELISA 和 Western blot 检测了来自西班牙安达卢西亚自治区的 145 只狗的血清,并将纯化的 Fe-SODe 与两种不同的抗原进行了比较:总寄生虫可溶性裂解物和未纯化的 Fe-SODe。为了验证使用纯化的 Fe-SODe 获得的结果,我们品尝了来自意大利伦巴第的 10 只感染婴儿利什曼原虫的狗的血清作为阳性对照。

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