A genoproteomic approach to detect peptide markers of bacterial respiratory pathogens

Honghui Wang, Steven K. Drake, Chen Yong, Marjan Gucek, Matthew A. Lyes, Avi Rosenberg, Erik Soderblom, M. Arthur Moseley, John P. Dekker, Anthony F. Suffredini

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Rapid identification of respiratory pathogens may facilitate targeted antimicrobial therapy. Direct identification of bacteria in bronchoalveolar lavage (BAL) by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry is confounded by interfering substances. We describe a method to identify unique peptide markers of 5 gramnegative bacteria by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for direct pathogen identification in BAL. METHODS: In silico translation and digestion were performed on 14-25 whole genomes representing strains of Acinetobacter baumannii, Moraxella catarrhalis, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Klebsiella pneumoniae. Peptides constituting theoretical core peptidomes in each were identified. Rapid tryptic digestion was performed; peptides were analyzed by LCMS/ MS and compared with the theoretical core peptidomes. High-confidence core peptides (false discovery rate 1%) were identified and analyzed with the lowest common ancestor search to yield potential speciesspecific peptide markers. The species specificity of each peptide was verified with protein BLAST. Further, 1 or 2 pathogens were serially diluted into pooled inflamed BAL, and a targeted LC-MS/MS assay was used to detect 25 peptides simultaneously. RESULTS: Five unique peptides with the highest abundance for each pathogen distinguished these pathogens with varied detection sensitivities. Peptide markers for A. baumannii and P. aeruginosa, when spiked simultaneously into inflamed BAL, were detected with as few as 3.6 (0.2) × 103 and 2.2 (0.6) × 103 colony-forming units, respectively, by targeted LC-MS/MS. CONCLUSIONS: This proof-of-concept study shows the feasibility of identifying unique peptides in BAL for 5 gram-negative bacterial pathogens, and it may provide a novel approach for rapid direct identification of bacterial pathogens in BAL.

Original languageEnglish (US)
Pages (from-to)1398-1408
Number of pages11
JournalClinical Chemistry
Volume63
Issue number8
DOIs
Publication statusPublished - Aug 1 2017
Externally publishedYes

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ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Wang, H., Drake, S. K., Yong, C., Gucek, M., Lyes, M. A., Rosenberg, A., ... Suffredini, A. F. (2017). A genoproteomic approach to detect peptide markers of bacterial respiratory pathogens. Clinical Chemistry, 63(8), 1398-1408. https://doi.org/10.1373/clinchem.2016.269647