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[婴儿利什曼原虫(利什曼原虫属)感染犬的蛋白质免疫印迹法、酶联免疫吸附测定和间接免疫荧光试验评估]

[Western blot, ELISA and indirect immunofluorescence test evaluation of Leishmania (Leishmania) infantum-infected dogs].

作者信息

Vargas-Duarte Jimmy J, López-Páez Myriam C, Escovar-Castro Jesús E, Fernández-Manrique José

机构信息

Instituto de Genética, Universidad Nacional de Colombia.

出版信息

Rev Salud Publica (Bogota). 2009 Aug;11(4):641-52. doi: 10.1590/s0124-00642009000400015.

Abstract

OBJECTIVE

Evaluating canine visceral leishmaniasis diagnostic test performance in Colombia and adapting the Western blot test in naturally and experimentally infected dogs.

METHODS

Sera were obtained from 10 experimentally L. Infantum-infected dogs, 5 naturally infected dogs, 16 healthy dogs, 26 Babesia canis, Erhlichia canis, Dirofilaria immitis, Trypanosoma cruzi and Leishmania (Viannia) spp infected dogs, 40 dogs from non-endemic areas and 150 from endemic areas. Sera were tested for L. infantum infection using immunofluorescent antibody (IFAT), ELISA and Western blot (WB) tests.

RESULTS

Positives results were obtained for 73 % of known infected dogs by the IFAT test and false positives were obtained for 2.5 % of non-infected dogs using WB. ELISA was not efficient for diagnosis. 24 antigenic fractions were recognised in tested sera using WB; however, 29, 34, 50, 69, 75, 86, 99 and 123 kDa bands were recognised in sera from dogs from non-endemic areas, healthy dogs and Trypanosoma cruzi, Erhlichia canis, Dirofilaria immitis and Babesia canis infected dogs. The 13 kDa fraction proved potentially useful for diagnosing canine visceral leishmaniasis.

CONCLUSIONS

The separate use of parasitological and serological test could lead to misdiagnosis of Leishmania infection; using both kinds of technique simultaneously is thus highly recommended.

摘要

目的

评估哥伦比亚犬内脏利什曼病诊断检测的性能,并对自然感染和实验感染犬的蛋白质印迹试验进行调整。

方法

采集了10只实验性感染婴儿利什曼原虫的犬、5只自然感染犬、16只健康犬、26只感染犬巴贝斯虫、犬埃立克体、犬恶丝虫、克氏锥虫和利什曼原虫(维扬亚属)的犬、40只来自非流行地区的犬和150只来自流行地区的犬的血清。使用免疫荧光抗体(IFAT)、酶联免疫吸附测定(ELISA)和蛋白质印迹(WB)试验检测血清中的婴儿利什曼原虫感染情况。

结果

IFAT试验在73%的已知感染犬中得到阳性结果,使用WB在2.5%的未感染犬中得到假阳性结果。ELISA对诊断无效。使用WB在检测血清中识别出24个抗原组分;然而,在来自非流行地区的犬、健康犬以及感染克氏锥虫、犬埃立克体、犬恶丝虫和犬巴贝斯虫的犬的血清中识别出了29、34、50、69、75、86、99和123 kDa条带。13 kDa组分被证明对诊断犬内脏利什曼病可能有用。

结论

单独使用寄生虫学和血清学检测可能导致利什曼原虫感染的误诊;因此强烈建议同时使用这两种技术。

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