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人体患者血清识别的曼氏迭宫绦虫裂头蚴抗原蛋白的二维免疫印迹分析

Two-dimensional immunoblot analysis of antigenic proteins of Spirometra plerocercoid recognized by human patient sera.

作者信息

Rahman Moizur, Lee Eung-Goo, Bae Young-An

机构信息

Department of Molecular Parasitology, Sungkyunkwan University School of Medicine and Center for Molecular Medicine, Samsung Biomedical Research Institute, Suwon 446-740,Republic of Korea.

出版信息

Parasitol Int. 2011 Jun;60(2):139-43. doi: 10.1016/j.parint.2011.01.002. Epub 2011 Jan 21.

Abstract

Sparganosis is caused by invasion of Spirometra plerocercoid into various tissues/organs. Subcutaneous sparganosis can be diagnosed and treated by worm removal, while visceral/cerebral sparganosis is not easy to diagnose. The diagnosis depends largely on the detection of specific antibodies circulating in the patients' sera. Previous studies demonstrated that 31 and 36kDa proteins of the sparganum invoked specific and sensitive antibody responses, but also showed cross reactions with cysticercosis sera. We enriched protein fractions containing 31-36kDa through gel filtration and examined immune recognition pattern against the patient sera by 2-dimensional electrophoresis (2-DE) followed by immunoblotting. Serum samples from sparganosis patients recognized 8-10 protein spots of 31 and 36kDa with different isoelectric point (pI) values with variable combinations, in which four spots of 31kDa with pIs 3.4, 3.9, 4.0 and 4.1, and one 36kDa spot (pI 3.5) appeared to be specifically reactive. One 31kDa protein spot with pI 3.3 and two spots of 36kDa with pIs 3.3 and 3.5 reacted crossly with neurocysticercosis sera. Neither sera from patients with other parasitic infections nor those from healthy controls showed positive reaction. Two-DE/immunoblot analysis might be highly available in differential serodiagnosis of human sparganosis.

摘要

裂头蚴病是由曼氏迭宫绦虫裂头蚴侵入各种组织/器官引起的。皮下裂头蚴病可通过取出虫体进行诊断和治疗,而内脏/脑裂头蚴病不易诊断。诊断很大程度上依赖于检测患者血清中循环的特异性抗体。先前的研究表明,裂头蚴的31kDa和36kDa蛋白可引发特异性和敏感的抗体反应,但也与囊尾蚴病血清出现交叉反应。我们通过凝胶过滤富集了含有31 - 36kDa的蛋白组分,并通过二维电泳(2-DE)随后进行免疫印迹来检测针对患者血清的免疫识别模式。裂头蚴病患者的血清样本识别出8 - 10个31kDa和36kDa的蛋白斑点,其等电点(pI)值不同且组合各异,其中4个pI分别为3.4、3.9、4.0和4.1的31kDa斑点以及1个pI为3.5的36kDa斑点似乎具有特异性反应。1个pI为3.3的31kDa蛋白斑点以及2个pI分别为3.3和3.5的36kDa斑点与神经囊尾蚴病血清发生交叉反应。其他寄生虫感染患者的血清以及健康对照者的血清均未显示阳性反应。二维电泳/免疫印迹分析在人体裂头蚴病的鉴别血清诊断中可能具有很高的实用性。

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