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: Nurulhasanah Othman, Zeehaida Mohamed, Maya Mazuwin Yahya, Voon Meng Leow, Boon Huat Lim, Rahmah Noordin
• Journal: Experimental parasitology (Exp Parasitol) Vol. 134 Issue 4 Pg. 504-10 (Aug 2013) ISSN: 1090-2449 [Electronic] United States
• PMID: 23680184 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Elsevier Inc. All rights reserved.

Chemical References

• Antibodies, Protozoan
• Antigens, Protozoan
• DNA, Protozoan
• Immunoglobulin G
• Protozoan Proteins

Topics

• Animals
• Antibodies, Protozoan (blood, immunology)
• Antigens, Protozoan (isolation & purification)
• Blotting, Western
• Cricetinae
• DNA, Protozoan (analysis)
• Electrophoresis, Polyacrylamide Gel
• Entamoeba histolytica (genetics, immunology)
• Enzyme-Linked Immunosorbent Assay
• Gas Chromatography-Mass Spectrometry
• Humans
• Immunoglobulin G (blood)
• Isoelectric Focusing
• Liver Abscess, Amebic (immunology, parasitology)
• Mesocricetus
• Polymerase Chain Reaction
• Proteomics
• Protozoan Proteins (genetics, immunology, isolation & purification)
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• Suppuration (immunology, parasitology)
• Tandem Mass Spectrometry