Rapid detection of colistin resistance protein MCR-1 by LC-MS/MS

Honghui Wang, Yong Chen, Jeffrey R. Strich, Steven K. Drake, Jung Ho Youn, Avi Z. Rosenberg, Marjan Gucek, Patrick T. McGann, Anthony F. Suffredini, John P. Dekker

Research output: Contribution to journalArticle

Abstract

Background: Colistin (polymyxin E) and polymixin B are important bactericidal antibiotics used in the treatment of serious infections caused by multi-drug resistant Gram-negative organisms. Transferrable plasmid-mediated colistin resistance, conferred by the product of the mcr-1 gene, has emerged as a global healthcare threat. Consequently, the rapid detection of the MCR-1 protein in clinical bacterial isolates has become increasingly important. We used a genoproteomic approach to identify unique peptides of the MCR-1 protein that could be detected rapidly by liquid chromatography tandem mass spectrometry (LC-MS/MS). Methods: MCR-1 tryptic peptides that were efficiently ionized and readily detectable were characterized in a set of mcr-1-containing isolates with triple quadrupole LC-MS. Three optimal peptides were selected for the development of a rapid multiple reaction monitoring LC-MS/MS assay for the MCR-1 protein. To investigate the feasibility of rapid detection of the MCR-1 protein in bacterial isolates using this assay, a blinded 99-sample test set was built that included three additional mcr-1-containing clinical isolates tested in triplicate (9 samples) and 90 negative control isolates. Results: All of the mcr-1-containing isolates in the test set were accurately identified with no false positive detections by three independent, blinded operators, yielding an overall performance of 100% sensitivity and specificity for multiple operators. Among the three peptides tested in this study, the best performing was DTFPQLAK. The isolate-to-result time for the assay as implemented is less than 90 min. Conclusions: This work demonstrates the feasibility of rapid detection of the MCR-1 protein in bacterial isolates by LC-MS/MS.

Original languageEnglish (US)
Article number8
JournalClinical Proteomics
Volume16
Issue number1
DOIs
StatePublished - Feb 26 2019

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Clinical Biochemistry

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    Wang, H., Chen, Y., Strich, J. R., Drake, S. K., Youn, J. H., Rosenberg, A. Z., Gucek, M., McGann, P. T., Suffredini, A. F., & Dekker, J. P. (2019). Rapid detection of colistin resistance protein MCR-1 by LC-MS/MS. Clinical Proteomics, 16(1), [8]. https://doi.org/10.1186/s12014-019-9228-2