Identification of rickettsial immunoreactive proteins using a proximity ligation assay Western blotting and the traditional immunoproteomic approach. (June 2018)

• Record Type: Journal Article
• Authors: Kowalczewska, Malgorzata; N'Djatchi, Aymar; Nappez, Claude; Alwassouf, Sulaf; Decloquement, Philippe; Armstrong, Nicholas; El Karkouri, Khalid; Edouard, Sophie; Raoult, Didier
• Is Part Of: Comparative immunology, microbiology and infectious diseases. Volume 58(2018)
• Page Start: 17
• Page End: 25
• Publication Date: 2018-06
• Subjects: ATBF African tick bite fever -- MSF Mediterranean spotted fever -- IFA immunofluorescence assay -- PCR polymerase chain reaction -- PLA WB proximity ligation assay-based Western blotting -- RCA rolling cycle amplification -- HRP horseradish peroxidase -- HBD healthy blood donors -- PCR polymerase chain reaction -- PLS-EDA partial least squares-enhanced discriminate analysis; Rickettsia africae -- Rickettsia conorii -- Serodiagnosis -- Immunoproteomics -- Proximity ligation assay; Communicable diseases in animals -- Periodicals; Veterinary immunology -- Periodicals; Veterinary microbiology -- Periodicals; Immunology -- Periodicals; Microbiology -- Periodicals; Communicable diseases -- Periodicals; Communicable Diseases -- immunology -- Periodicals; Communicable Diseases -- veterinary -- Periodicals; Allergy and Immunology -- Periodicals; Microbiology -- Periodicals; Veterinary Medicine -- Periodicals; Immunologie -- Périodiques; Microbiologie -- Périodiques; Maladies infectieuses -- Périodiques; Communicable diseases; Immunology; Microbiology; Electronic journals; Periodicals; 636.08969
• DOI: 10.1016/j.cimid.2018.06.004
• Languages: English
• ISSNs: 0147-9571

Abstract

Graphical abstract: Highlights: Two proteomics- based techniques are used for screening R. africae and R. conorii new diagnostic protein targets. New technique named in situ proximity ligation assay in 2-D Western blotting has been developed. Both approaches were compared and evaluated. Several rickettsial proteins with diagnostic potential were identified. They may be proposed for developement of rickettsial serodiagnosis. 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.