Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi 221 005, India.
Parasitol Res. 2012 Oct;111(4):1851-5. doi: 10.1007/s00436-012-2931-9. Epub 2012 May 5.
The present study was aimed to detect urinary proteins excreted in kala-azar patients. Urinary proteins were isolated by ammonium sulfate precipitation and purified by Amicon ultra using 3 kDa cutoff membrane device. The proteins were resolved on 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis and blots were developed with the patients sera. The excretion of urinary proteins was differential in kala-azar patients. A total eight proteins of molecular weights 25, 28, 46, 54, 57, 60, 72, and 95 kDa were detected in the urine of visceral leishmaniasis (VL) patients. On blot, the 57 kDa protein was found to be host origin and characterized as human kininogen. All other proteins were leishmanial origin. Out of 50 urine samples analyzed, the kininogen was detected in 45 urine samples. Following treatment, this protein was not detectable in the urine samples of any patient. The appearance of kininogen in urine of VL patients offers a novel possibility for the development of diagnostic tool and a test of cure.
本研究旨在检测黑热病患者尿液中排出的蛋白质。通过硫酸铵沉淀法分离尿液蛋白，并使用 cutoff 值为 3 kDa 的 Amicon ultra 超滤设备进行纯化。将蛋白质在 12%十二烷基硫酸钠聚丙烯酰胺凝胶电泳上进行分离，并用患者血清进行印迹分析。黑热病患者尿液中的蛋白质排泄存在差异。在内脏利什曼病（VL）患者的尿液中检测到分子量为 25、28、46、54、57、60、72 和 95 kDa 的 8 种蛋白质。在印迹中，发现 57 kDa 蛋白为宿主来源，并鉴定为人激肽原。所有其他蛋白质均为利什曼原虫来源。在分析的 50 个尿液样本中，45 个尿液样本中检测到激肽原。治疗后，该蛋白在任何患者的尿液样本中均无法检测到。激肽原在 VL 患者尿液中的出现为开发诊断工具和治疗检测提供了新的可能性。
