Rapid identification of New Delhi metallo-β-lactamase (NDM) using tryptic peptides and LC-MS/MS

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

Research output: Contribution to journalArticle

Abstract

There is significant interest in the development of mass spectrometry (MS) methods for antimicrobial resistance protein detection, given the ability of these methods to confirm protein expression. In this work, we studied the performance of a liquid chromatography, tandem MS multiple-reaction monitoring (LC-MS/MS MRM) method for the direct detection of the New Delhi metallo-lactamase (NDM) carbapenemase in clinical isolates. Using a genoproteomic approach, we selected three unique peptides (SLGNLGDADTEHYAASAR, AFGAAFPK, and ASMIVMSHS APDSR) specific to NDM that were efficiently ionized and spectrally well-defined. These three peptides were used to build an assay with turnaround time of 90 min. In a blind set, the assay detected 21/24 blaNDM-containing isolates and 76/76 isolates with negative results, corresponding to a sensitivity value of 87.5% (95% confidence interval [CI], 67.6% to 97.3%) and a specificity value of 100% (95% CI, 95.3% to 100%). One of the missed identifications was determined by protein fractionation to be due to low (0.1 fm/g) NDM protein expression (below the assay limit of detection). Parallel disk diffusion susceptibility testing demonstrated this isolate to be meropenem susceptible, consistent with low NDM expression. Total proteomic analysis of the other two missed identifications did not detect NDM peptides but detected other proteins expressed from the blaNDM-containing plasmids, confirming that the plasmids were not lost. The measurement of relative NDM concentrations over the entire isolate test set demonstrated variability spanning 4 orders of magnitude, further confirming the remarkable range that may be seen in levels of NDM expression. This report highlights the sensitivity of LC-MS/MS to variations in NDM protein expression, with implications for how this technology may be used.

Original languageEnglish (US)
Article numbere00461-19
JournalAntimicrobial agents and chemotherapy
Volume63
Issue number9
DOIs
StatePublished - Jan 1 2019

Keywords

  • Mass spectrometry
  • Multiple-reaction monitoring
  • New Delhi metallo-lactamase
  • Tryptic peptide

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

Fingerprint Dive into the research topics of 'Rapid identification of New Delhi metallo-β-lactamase (NDM) using tryptic peptides and LC-MS/MS'. Together they form a unique fingerprint.

  • Cite this

    Wang, H., Strich, J. R., Drake, S. K., Chen, Y., Youn, J. H., Rosenberg, A., Gucek, M., Dekker, J. P., & Suffredini, A. F. (2019). Rapid identification of New Delhi metallo-β-lactamase (NDM) using tryptic peptides and LC-MS/MS. Antimicrobial agents and chemotherapy, 63(9), [e00461-19]. https://doi.org/10.1128/AAC.00461-19