Institute for Research in Molecular Medicine, Universiti Sains Malaysia USM, Penang, Malaysia.
Exp Parasitol. 2013 Aug;134(4):504-10. doi: 10.1016/j.exppara.2013.05.001. Epub 2013 May 13.
Entamoeba histolytica is a causative agent of amoebic liver abscess (ALA) and is endemic in many underdeveloped countries. We investigated antigenic E. histolytica proteins in liver abscess aspirates using proteomics approach. Pus samples were first tested by real-time PCR to confirm the presence of E. histolytica DNA and the corresponding serum samples tested for E. histolytica-specific IgG by a commercial ELISA. Proteins were extracted from three and one pool(s) of pus samples from ALA and PLA (pyogenic liver abscess) patients respectively, followed by analysis using isoelectric focussing, SDS-PAGE and Western blot. Unpurified pooled serum samples from infected hamsters and pooled human amoebic-specific IgG were used as primary antibodies. The antigenic protein band was excised from the gel, digested and analysed by MALDI-TOF/TOF and LC-MS/MS. The results using both primary antibodies showed an antigenic protein band of ∼14kDa. Based on the mass spectrum analysis, putative tyrosine kinase is the most probable identification of the antigenic band.
溶组织内阿米巴是肝脓肿（ALA）的病原体，在许多欠发达国家流行。我们使用蛋白质组学方法研究了肝脓肿抽吸物中的抗原性溶组织内阿米巴蛋白。脓液样本首先通过实时 PCR 检测以确认溶组织内阿米巴 DNA 的存在，相应的血清样本通过商业 ELISA 检测溶组织内阿米巴特异性 IgG。从 ALA 和 PLA（化脓性肝脓肿）患者的三个和一个脓液样本池分别提取蛋白质，然后进行等电聚焦、SDS-PAGE 和 Western blot 分析。未纯化的感染仓鼠和人阿米巴特异性 IgG 混合血清样本用作初级抗体。从凝胶中切下抗原性蛋白带，进行消化，并通过 MALDI-TOF/TOF 和 LC-MS/MS 进行分析。两种初级抗体的结果均显示出约 14kDa 的抗原性蛋白带。基于质谱分析，酪氨酸激酶是抗原带的最可能鉴定。
