Multiplex PCR for detection of acquired carbapenemase genes

Poirel, Laurent; Walsh, Timothy R.; Cuvillier, Vincent; Nordmann, Patrice

doi:10.1016/j.diagmicrobio.2010.12.002

Cited by 1,475 publications

(795 citation statements)

References 26 publications

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“…PCR for bla-TEM , bla-SHV , and bla-CTX-M was carried out as previously described by Jemima et al 10 PCR amplification with some modifications in the annealing temperatures for bla OXA-2 , bla OXA-10 , bla OXA-48 , bla VIM , bla NDM , bla GES , bla VEB , bla IMP , and bla PER was performed as previously described by Shacheraghi et al 11 and Poirel et al 12 The annealing temperatures and primers are described in Table 1. PCR products were visualised by electrophoresis in 1.5% agarose gel for 40 min at 120 V then sequenced using Sanger Dideoxy sequencing technology (Inqaba Biotech, Pretoria, South Africa).…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

Molecular characterisation of multidrug-resistant Pseudomonas aeruginosa from a private hospital in Durban, South Africa

Adjei

Govinden

Moodley

et al. 2017

Southern African Journal of Infectious Diseases

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Background: Multi-drug resistant Pseudomonas aeruginosa pose a clinical challenge globally. This study delineated the molecular mechanisms of resistance to β-lactam antibiotics in multidrug-resistant P. aeruginosa isolated from a single private hospital in Durban, South Africa and ascertained clonality with regard to the isolates carrying β-lactamase genes. Methods: Seventeen P. aeruginosa isolates recovered from sputum, urine, catheter tips, pus swabs, nasal swabs and endotracheal aspirates underwent MIC determination, and phenotypic screening using the Double Disk Synergy Test (DDST) and Modified Hodge Test (MHT) to identify putative extended-spectrum β-lactamases (ESBLs), metallo-β-lactamases and other carbapenemases. Selected β-lactamase encoding genes were genotypically confirmed by PCR and sequencing. REP-PCR was conducted to determine the clonal relatedness of the 11 isolates carrying β-lactamase genes. Results: Sixteen isolates (94%) were resistant to aztreonam and piperacillin, 15 isolates (88%) were resistant to imipenem and ticarcillin, 14 (82%) were resistant to meropenem, and 13 isolates (76%) were resistant to ceftazidime and piperacillin/tazobactam. Resistance to ciprofloxacin and amikacin were 82% and 29% respectively. Of the 17 isolates tested, GES-2, VIM-2 and OXA-21 were present in 10 (59%) four (24%) and one (6%) of the isolates respectively. Three of the isolates harboured both GES-2 and VIM-2 and one isolate harboured OXA-21 and VIM-2. REP-PCR revealed seven clusters with clusters A and F having two (18%) and four (36%) isolates respectively, while the remaining five isolates were unrelated. Conclusion: GES-2 and VIM-2 enzymes were predominantly responsible for carbapenemase resistance. Clones A and F intimated patient-to-patient spread within the ICU and surgical ICU. This apparent dissemination as well as the multi-drug resistance observed points to sub-optimal infection prevention and control and dwindling antibiotic treatment options for P. aeruginosa respectively in this institution.

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Section: Polymerase Chain Reactionmentioning

confidence: 99%

Molecular characterisation of multidrug-resistant Pseudomonas aeruginosa from a private hospital in Durban, South Africa

Adjei

Govinden

Moodley

et al. 2017

Southern African Journal of Infectious Diseases

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“…Se realizó mediante los métodos de Hodge modificado 19 , sinergia con ácido fenil borónico (AFB) 21 23 . Los productos de amplificación se sometieron a electroforesis en gel de agarosa al 2%, luego se colorearon con bromuro de etidio (0,5 μg.ml -1 ) en tampón TBE 1X, durante 30 min a 10 V.cm -1 .…”

Section: Detección De Carbapenemasas Por Métodos Fenotípicosunclassified

Relación clonal y detección del gen blaKPC en cepas de Klebsiella pneumoniae resistentes a carbapenémicos, en un hospital de Venezuela

Martínez¹,

Araque

Roduifo³

et al. 2016

Rev. chil. infectol.

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“…DNA in supernatant was used as a template under standard PCR procedure [9] with a series of different sets of primers. Detection of the known acquired CHDLs (bla OxA-23, bla , and bla OxA-58 ) [14], class A extended-spectrum b-lactamases (bla PER , bla VEB , and bla GES ) [14,15], MBL (bla IMP , bla VIM , bla SIM , bla NDM ) [16,17] and the intrinsic bla OxA-51-like gene was performed as previously described [14].…”

Section: Pcr Amplification and Plasmid Typingmentioning

confidence: 99%

Multidrug-resistant Acinetobacter baumannii strains carrying the blaOxA-23 and the blaGES-11 genes in a neonatology center in Tunisia

Charfi

Mansour²,

Khalifa³

et al. 2014

Microbial Pathogenesis

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Multidrug-resistant and difficult-to-treat Acinetobacter baumannii may be responsible for nosocomial infections. The production of carbapenem-hydrolyzing class D b-lactamases (CHDLs) and extendedspectrum b-lactamase (ESBLs) of the GES type possessing a carbapenemase activity has been increasingly reported worldwide in A. baumannii. The aim of this study was to analyze the resistance mechanisms of two carbapenem resistant A. baumannii clinical isolates recovered in a neonatology center in the center-east of Tunisia.Two carbapenem resistant A. baumannii isolates were recovered. The first isolate co-harbored the bla GES-11 ESBL gene and the bla OxA-23 CHDL gene. Analyses of the genetic location indicated that the bla GES-11 gene was plasmid located (Gr6). However, the bla OxA-23 gene was located on the chromosome. The second strain had only the bla OxA-23 CHDL gene, which was plasmid located.This study showed the first description of the GES-type b-lactamase in A. baumannii in Tunisia.

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Multiplex PCR for detection of acquired carbapenemase genes

Cited by 1,475 publications

References 26 publications

Molecular characterisation of multidrug-resistant Pseudomonas aeruginosa from a private hospital in Durban, South Africa

Molecular characterisation of multidrug-resistant Pseudomonas aeruginosa from a private hospital in Durban, South Africa

Relación clonal y detección del gen blaKPC en cepas de Klebsiella pneumoniae resistentes a carbapenémicos, en un hospital de Venezuela

Multidrug-resistant Acinetobacter baumannii strains carrying the blaOxA-23 and the blaGES-11 genes in a neonatology center in Tunisia

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