title: Entamoeba histolytica antigenic protein detected in pus aspirates from patients with amoebic liver abscess.
abstract: 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.
authors.affiliation: Institute for Research in Molecular Medicine, Universiti Sains Malaysia USM, Penang, Malaysia.
authors.forename: Nurulhasanah
authors.initials: N
authors.lastname: Othman
authors.forename: Zeehaida
authors.initials: Z
authors.lastname: Mohamed
authors.forename: Maya Mazuwin
authors.initials: MM
authors.lastname: Yahya
authors.forename: Voon Meng
authors.initials: VM
authors.lastname: Leow
authors.forename: Boon Huat
authors.initials: BH
authors.lastname: Lim
authors.forename: Rahmah
authors.initials: R
authors.lastname: Noordin
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood / Q000276:immunology; D000953:Antigens, Protozoan / Q000302:isolation & purification*; D015153:Blotting, Western; D006224:Cricetinae; D016054:DNA, Protozoan / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D004748:Entamoeba histolytica / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007525:Isoelectric Focusing; D008101:Liver Abscess, Amebic / Q000276:immunology* / Q000469:parasitology; D008647:Mesocricetus; D016133:Polymerase Chain Reaction; D040901:Proteomics; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013492:Suppuration / Q000276:immunology / Q000469:parasitology; D053719:Tandem Mass Spectrometry
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D007074:Immunoglobulin G; D015800:Protozoan Proteins
keywords: Amoebic liver abscess (ALA); Entamoeba histolytica; MS/MS spectrometry; Two-dimensional electrophoresis (2-DE)