Saldaña A, Orn A, Henriksson J, Sousa O E
Centro de Investigacion y Diagnóstico de Enfermedades Parasitarias (CIDEP), Facultad de Medicina, Universidad de Panamá.
Rev Med Panama. 1993 Jan;18(1):41-52.
In American man can be infected with two trypanosomes: Trypanosoma cruzi, the etiological agent of Chagas' disease and Trypanosoma rangeli, a suspected nonpathogenic parasite. In this communication are presented 4 methods in order to improve the current knowledge about the specific identification of these parasites. Using the SDS-PAGE technique it was possible to differentiate between T. rangeli. and T. cruzi based in at less 4 protein bands with a relative molecular weights of 93, 77-73, 63 and 54-52 KDa. These polypeptides were found only in T. rangeli electrophoretic profiles. An ELISA test showed that the antigenic composition found in the enzyme cisteine proteinase (cruzipain) is specific for T. cruzi epimastigotes. Antigenic analysis by Western blot assay, proved that T. rangeli and not T. cruzi present antigenic bands with a Mr of 142, 63, 54, 51, 49, 43, 39 and 24 KDa. Finally, using the Southern blot procedure, it was confirmed that SAPA, a DNA sequence originally identified in the T. cruzi, genome, is absent in T. rangeli nuclear DNA. These initial observations revealed that it is possible to identify both parasites using the described methods, however further works are required to clarify the biochemical, immunological and molecular relationship between T. rangeli and T. cruzi.
在美国,男性可能感染两种锥虫:恰加斯病的病原体克氏锥虫和一种疑似无致病性的寄生虫兰氏锥虫。本文介绍了4种方法,以增进目前对这些寄生虫特异性鉴定的了解。使用SDS-PAGE技术,基于至少4条相对分子量为93、77 - 73、63和54 - 52 kDa的蛋白带,有可能区分兰氏锥虫和克氏锥虫。这些多肽仅在兰氏锥虫的电泳图谱中发现。一项ELISA试验表明,在酶半胱氨酸蛋白酶(克氏锥虫蛋白酶)中发现的抗原成分对克氏锥虫前鞭毛体具有特异性。通过蛋白质印迹分析进行的抗原分析证明,兰氏锥虫而非克氏锥虫呈现出分子量为142、63、54、51、49、43、39和24 kDa的抗原条带。最后,使用Southern印迹法,证实了最初在克氏锥虫基因组中鉴定出的DNA序列SAPA在兰氏锥虫核DNA中不存在。这些初步观察结果表明,使用所述方法可以鉴定这两种寄生虫,然而,需要进一步的研究来阐明兰氏锥虫和克氏锥虫之间的生化、免疫和分子关系。