Identification of rickettsial immunoreactive proteins using a proximity ligation assay Western blotting and the traditional immunoproteomic approach

Comp Immunol Microbiol Infect Dis. 2018 Jun:58:17-25. doi: 10.1016/j.cimid.2018.06.004. Epub 2018 Jun 15.

Abstract

The closely related species Rickettsia conorii and R. africae are both etiological agents of rickettsiosis, a tick-borne serious infective disease. The laboratory diagnosis is based on serology, but remains not enough specific to provide the diagnosis at the species level. Here, we attempted to identify specific proteins that would enable the discrimination of R. africae sp from R. conorii sp infections. We screened 22 R. africae- and 24 R. conorii-infected sera at different course of infection using a traditional immunoproteomic approach. In parallel, we focused on the technical development of a "relatively new technique" named a proximity ligation assay coupled to two-dimensional Western blotting. The top range markers of R. africae early infection were rpoA, atpD, and acnA, ORF0029, R. africae active infection were rOmpB β-peptide, OmpA, groEL and ORF1174, early R. conorii infection was prsA, RC0031, pepA, R. conorii active infection were ftsZ, cycM and rpoA. They are candidates for serodiagnosis of rickettsioses.

Keywords: Immunoproteomics; Proximity ligation assay; Rickettsia africae; Rickettsia conorii; Serodiagnosis.

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood*
  • Bacterial Proteins / immunology
  • Bacterial Proteins / isolation & purification
  • Biomarkers / blood
  • Blotting, Western*
  • France / epidemiology
  • Humans
  • Proteomics*
  • Rickettsia / chemistry
  • Rickettsia / genetics
  • Rickettsia / immunology*
  • Rickettsia Infections / blood
  • Rickettsia Infections / diagnosis*
  • Rickettsia Infections / epidemiology
  • Rickettsia Infections / immunology
  • Rickettsia conorii / chemistry
  • Rickettsia conorii / genetics
  • Rickettsia conorii / immunology
  • Serologic Tests / methods
  • Ticks / microbiology

Substances

  • Antibodies, Bacterial
  • Bacterial Proteins
  • Biomarkers