Leishmania infantum secreted iron superoxide dismutase purification and its application to the diagnosis of canine Leishmaniasis |
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Authors: | Silvia S. Longoni,Manuel Sá nchez-MorenoJorge Emilio Rivera Ló pez,Clotilde Marí n |
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Affiliation: | Departamento de Parasitología, Facultad de Ciencias, Universidad de Granada, Severo Ochoa s/n, 18071 Granada, Spain |
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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. |
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Keywords: | Leishmania infantum Ion exchange chromatography Molecular sieve chromatography Iron superoxide dismutase Canine Leishmaniasis Spain ELISA Western blot Oxidative stress Protein purification |
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