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High-performance method to detection of Klebsiella pneumoniae Carbapenemase in Enterobacterales by LC-MS/MS.

Authors
  • Lovison, Otávio A1, 2
  • Rau, Renata B1, 2, 3
  • Lima-Morales, Daiana1
  • Almeida, Evellyn K1, 4
  • Crispim, Marina N1
  • Barreto, Fabiano3
  • Barth, Afonso L1, 2, 4
  • Martins, Andreza F5, 6
  • 1 Laboratório de Pesquisa em Resistência Bacteriana (LABRESIS), Centro de Pesquisa Experimental, Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, Brazil. , (Brazil)
  • 2 Programa de Pós-Graduação em Ciências Farmacêuticas (PPGCF), Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil. , (Brazil)
  • 3 Laboratório Nacional Agropecuário no Rio Grande do Sul (LANAGRO/RS), Porto Alegre, Brazil. , (Brazil)
  • 4 Faculdade de Farmácia - Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil. , (Brazil)
  • 5 Programa de Pós-Graduação em Ciências Farmacêuticas (PPGCF), Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil. [email protected] , (Brazil)
  • 6 Laboratório de Microbiologia Aplicada, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil. [email protected] , (Brazil)
Type
Published Article
Journal
Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]
Publication Date
Sep 01, 2020
Volume
51
Issue
3
Pages
1029–1035
Identifiers
DOI: 10.1007/s42770-019-00222-y
PMID: 31989451
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Carbapenem-resistant Enterobacterales (CREs) have been recognized as an important threat to global health. CRE cause the majority of the difficult-to-treat infections in health-care settings and are associated with high mortality. Klebsiella pneumoniae carbapenemase (KPC)-producing CREs, in particular Klebsiella pneumoniae, are globally disseminated and responsible for a large number of outbreaks. Development of rapid methods for KPC detection can provide great clinical and epidemiological benefits to prevent KPC dissemination. The aim of this study was to standardize and validate a LC-MS/MS method to detect KPC. This method was also tested against a broad variety of species, including CRE with other carbapenemase genes and the recently reported mcr-1. For validation, 111 isolates with reduced susceptibility to carbapenems were selected (49 KPC-positive and 62 KPC-negative). The presence of four tryptic peptides related to the KPC enzyme was evaluated, and the identification of at least two of them classified the isolate as "KPC-positive." The LTLGSALAAPQR and LALEGLGVNGQ peptides were both detected in 47 of 49 isolates with the blaKPC gene. The other two peptides, GFLAAAVLAR and APIVLAVYTR, were detected in 46 and 19 isolates with the blaKPC gene, respectively. The method correctly classified 47 of 49 KPC-positive and all KPC-negative isolates yielding 96.07% of sensitivity and 100% of specificity. In conclusion, our results demonstrate that the KPC peptide markers were robustly detected by the method which presented high sensitivity and full specificity and therefore can be used as a reliable method to identify this resistance mechanism.

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