Rapid Identification of New Delhi Metallo-β-Lactamase (NDM) Using Tryptic Peptides and LC-MS/MS
| العنوان: | Rapid Identification of New Delhi Metallo-β-Lactamase (NDM) Using Tryptic Peptides and LC-MS/MS |
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| المؤلفون: | Jung Ho Youn, Honghui Wang, Avi Z. Rosenberg, Marjan Gucek, John P. Dekker, Steven K. Drake, Yong Chen, Jeffrey R Strich, Anthony F. Suffredini |
| المصدر: | Antimicrobial Agents and Chemotherapy. 63 |
| بيانات النشر: | American Society for Microbiology, 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | Microbial Sensitivity Tests, beta-Lactams, Peptide Mapping, Meropenem, beta-Lactam Resistance, beta-Lactamases, 03 medical and health sciences, Plasmid, Tandem Mass Spectrometry, Lc ms ms, medicine, Trypsin, Pharmacology (medical), Amino Acid Sequence, 030304 developmental biology, Pharmacology, Detection limit, Analytical Procedures, 0303 health sciences, 030306 microbiology, Chemistry, Selected reaction monitoring, Tryptic peptide, Gene Expression Regulation, Bacterial, Molecular biology, Orders of magnitude (mass), Anti-Bacterial Agents, Isoenzymes, Klebsiella pneumoniae, Infectious Diseases, Proteolysis, Biological Assay, New delhi, Peptides, Chromatography, Liquid, Plasmids, medicine.drug |
| الوصف: | 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-β-lactamase (NDM) carbapenemase in clinical isolates. Using a genoproteomic approach, we selected three unique peptides (SLGNLGDADTEHYAASAR, AFGAAFPK, and ASMIVMSHSAPDSR) 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 bla(NDM)-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/μg) 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 bla(NDM)-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. |
| تدمد: | 1098-6596 0066-4804 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1cb2a838798192158bf105214f06eac9 https://doi.org/10.1128/aac.00461-19 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....1cb2a838798192158bf105214f06eac9 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10986596 00664804 |
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