Reliable tool for detection of novel Coxiella burnetii antigens, using immobilized human polyclonal antibodies

J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Mar 15:1047:84-91. doi: 10.1016/j.jchromb.2016.08.044. Epub 2016 Aug 31.

Abstract

Coxiella burnetii (C. burnetii) is the etiological agent of a Q fever-the re-emerging disease with considerable economic impact. Due to many similar symptoms with commonly occurring infections, its clinical diagnosis is very difficult. Thus, a strong effort should be taken to raise the awareness and develop a robust strategy for an accurate diagnosis. The identification of specific C. burnetii biomarkers could be valuable for a sensitive and selective diagnosis of the disease. Herein, we described a workflow to identify immunoreactive proteins of C. burnetii with a high confidence. It is based on immunocapturing of bacterial antigens by biofunctionalized magnetic microspheres, followed by tandem mass spectrometry (MS/MS) identification. We detected dozens of previously reported antigens and proposed 15 novel biomarkers, which specificity was confirmed by in silico epitope analysis. Among them, the cardiolipin synthetase participating in the synthesis of cardiolipin was recognized. This biomarker could play a critical role in the early management of acute Q fever and prevention of Q fever endocarditis.

Keywords: Biofunctionalized magnetic beads; Biomarkers; Immunoreactive proteins; LC–MS/MS; Q fever; Serological proteome analysis (SERPA).

MeSH terms

  • Antibodies, Bacterial / blood
  • Antibodies, Bacterial / chemistry
  • Antibodies, Bacterial / immunology*
  • Antibodies, Immobilized / blood
  • Antibodies, Immobilized / chemistry
  • Antibodies, Immobilized / immunology
  • Antigens, Bacterial / analysis
  • Antigens, Bacterial / blood
  • Antigens, Bacterial / immunology*
  • Bacterial Proteins / analysis
  • Bacterial Proteins / blood
  • Bacterial Proteins / immunology*
  • Chromatography, Liquid
  • Coxiella burnetii / immunology*
  • Humans
  • Proteomics
  • Q Fever / blood
  • Q Fever / diagnosis*
  • Q Fever / immunology
  • Tandem Mass Spectrometry

Substances

  • Antibodies, Bacterial
  • Antibodies, Immobilized
  • Antigens, Bacterial
  • Bacterial Proteins