MSACL 2019 EU Abstract

Discovery of Species-Unique Peptides as Candidate Peptide Biomarkers for Respiratory Tract Pathogens Using Tandem Mass Spectrometry-based Proteotyping

Roger Karlsson(1,2,4,6), Annika Thorsell(5), Margarita Gomila(7), Francisco Salvà-Serra(1,2,4,7), Hedvig Engström-Jakobsson(1,4), Lucia Gonzales-Siles(1,2,4), Daniel Jaén-Luchoro(1,2,4), Susann Skovbjerg(1,2), Johannes Fuchs(5), Anders Karlsson(6), Fredrik Boulund(8,9), Anna Johnning(4,9), Erik Kristiansson(4,9), Edward R.B. Moore(1,2,3,4 )
(1) Dept of Clinical Microbiology, Sahlgrenska University Hospital, Region Västra Götaland, Sweden (2) Dept of Infectious Diseases, Sahlgrenska Academy of the University of Gothenburg, Gothenburg, Sweden (3) Culture Collection University of Gothenburg (CCUG), Sahlgrenska Academy of the University of Gothenburg, Gothenburg, Sweden (4) Centre for Antibiotic Resistance Research (CARe), University of Gothenburg, Gothenburg, Sweden (5) Proteomics Core Facility at Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden (6) Nanoxis Consulting AB, Gothenburg, Sweden (7) Microbiology, Dept of Biology, University of the Balearic Islands, Palma de Mallorca, Spain (8) Center for Translational Microbiome Research (CTMR), Dept of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden (9) Dept of Mathematical Sciences, Chalmers, Gothenburg, Sweden

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Roger Karlsson (Presenter) Sahlgrenska University Hospital

Relevant Financial Disclosures (within past 24 months) Salary Nanoxis Consulting AB Discussing Products? NO, presenter will NOT present educational content related to products/services of the ineligible company(ies) listed above. Conflict mitigation NOT REQUIRED.

INTRODUCTION. Respiratory tract infections are a major cause of clinical treatment, but clinical assessments and standard clinical laboratory protocols are time consuming and often insufficient for reliable diagnoses. New methods, preferably applied directly to clinical samples, without any culturing, are needed to improve diagnostics, provide adequate treatment and reduce the use of antibiotics and associated development of antibiotic resistance.
OBJECTIVES. To use tandem mass spectrometry-based proteotyping for discovery of species-unique peptides as potential biomarkers for bacterial pathogens commonly found in respiratory tract infections: Staphylococcus aureus; Moraxella catarrhalis; Haemophilus influenzae and Streptococcus pneumoniae.
METHODS. The workflow combined high-resolution accurate-mass (HRAM) tandem MS and the TCUP bioinformatics pipeline in the search for new species-unique peptides as potential biomarkers for respiratory pathogens. Species-unique peptides were discovered by analyzing both pure culture bacterial reference strains, which reflect the genomic variation in the four species and also clinical respiratory samples, without prior culturing, thus highlighting proteins expressed in clinical infection conditions.
RESULTS. For each of the four respiratory tract infectious pathogens, 15-20 peptide biomarker candidates most prominently found in direct analyses of clinical samples, were identified. To demonstrate the potential of using these species-unique peptides as effective peptide biomarker candidates for diagnosis of infectious diseases, targeted tandem MS analyses of clinical samples was performed and the relevance of the peptide biomarkers for identifying the pathogens, i.e., proteotyping, was shown.