Detection of expanded-spectrum β-lactamases in Gram-negative bacteria in the 21st century

Al-Bayssari, Charbel; Dabboussi, Fouad; Hamzé, Monzer; Rolain, Jean‐Marc

doi:10.1586/14787210.2015.1066247

Cited by 20 publications

(11 citation statements)

References 199 publications

(158 reference statements)

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“…As screening for multidrug-resistant colonizers is still poorly standardized [ 10 ], it remains questionable what a negative screening result after previous colonization with resistant bacteria really means: either definite vanishing of the resistant pathogen or just a shift to a concentration below the detection limit. As preanalytic conditions like swabbing techniques [ 11 ] and the use of enrichment broths [ 12 ] were shown to relevantly affect the reliability of enteric screening approaches, it is highly likely that a proportion of individuals with apparently cleared colonization remains colonized with multidrug-resistant bacteria on a level below the diagnostic threshold.…”

Section: Introductionmentioning

confidence: 99%

Resistant Gram-Negative Bacteria and Diagnostic Point-of-Care Options for the Field Setting during Military Operations

Frickmann

Podbielski²,

Kreikemeyer³

2018

BioMed Research International

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The spread of multidrug-resistant bacteria in resource-poor settings affects the military medical service in case of deployments of soldiers to war and crisis zones. Patients with war injuries are prone to colonization or infection with multidrug-resistant bacteria. Resistant Gram-negative bacteria play a dominant role in military wound infections. Problematic hygiene conditions on deployment facilitate exposition of soldiers with subsequent colonization. Although colonizing strains are frequently cleared from their hosts after returning from deployment, transmission to close contacts of the soldiers in the home country cannot be excluded and therapeutic options are reduced if colonization progresses to invasive infection. Since sophisticated culture-based diagnostic approaches are typically not available in the field setting on deployment, molecular rapid diagnostic test systems are an option for transmission control if the locally prevalent molecular resistance mechanisms are known. Efforts for global resistance surveillance can contribute to better understanding of resistance distribution and spread at deployment sites. This review summarizes experience of the military medical services with multidrug resistance on deployment and with the influx of resistant strains to the home country and discusses potential use of available molecular rapid test systems as an option for the field setting.

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Section: Introductionmentioning

confidence: 99%

Resistant Gram-Negative Bacteria and Diagnostic Point-of-Care Options for the Field Setting during Military Operations

Frickmann

Podbielski²,

Kreikemeyer³

2018

BioMed Research International

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“…While swabbing techniques and the kind of used swabs affect the sensitivity of screening for MRSA (Bartolitius et al, 2014; Warnke et al, 2014a,b,c), even basal questions like the need for broth enrichment (Murk et al, 2009; Jazmati et al, 2016), required assessment strategies, and the number of samples (Ho et al, 2012; Vrioni et al, 2012; Rybczynska et al, 2014; Al-Bayssari et al, 2015) are under debate for screening for colonizers of the gut like vancomycin-resistant Enterococcus spp. or beta-lactam-resistant Enterobacteriaceae.…”

Section: Methodical Issuesmentioning

confidence: 99%

More Pathogenicity or Just More Pathogens?—On the Interpretation Problem of Multiple Pathogen Detections with Diagnostic Multiplex Assays

et al. 2017

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Modern molecular diagnostic approaches in the diagnostic microbiological laboratory like real-time quantitative polymerase chain reaction (qPCR) have led to a considerable increase of diagnostic sensitivity. They usually outperform the diagnostic sensitivity of culture-based approaches. Culture-based diagnostics were found to be insufficiently sensitive for the assessment of the composition of biofilms in chronic wounds and poorly standardized for screenings for enteric colonization with multi-drug resistant bacteria. However, the increased sensitivity of qPCR causes interpretative challenges regarding the attribution of etiological relevance to individual pathogen species in case of multiple detections of facultative pathogenic microorganisms in primarily non-sterile sample materials. This is particularly the case in high-endemicity settings, where continuous exposition to respective microorganisms leads to immunological adaptation and semi-resistance while considerable disease would result in case of exposition of a non-adapted population. While biofilms in chronic wounds show higher pathogenic potential in case of multi-species composition, detection of multiple pathogens in respiratory samples is much more difficult to interpret and asymptomatic enteric colonization with facultative pathogenic microorganisms is frequently observed in high endemicity settings. For respiratory samples and stool samples, cycle-threshold-value-based semi-quantitative interpretation of qPCR results has been suggested. Etiological relevance is assumed if cycle-threshold values are low, suggesting high pathogen loads. Although the procedure is challenged by lacking standardization and methodical issues, first evaluations have led to promising results. Future studies should aim at generally acceptable quantitative cut-off values to allow discrimination of asymptomatic colonization from clinically relevant infection.

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“…Less than 20 years after their first identification, these microorganisms already represented one of the most important groups of nosocomial pathogens (Gniadkowski, 2001). Today, extended-spectrum β-lactamases (ESBL) are the most common resistance mechanism of Gram-negative bacteria against β-lactam antibiotics (Al-Bayssari et al, 2015) and have become a concern for public health, with growing infection and colonization rates worldwide (Karanika et al, 2016; McDanel et al, 2017). ESBL-producing bacteria have also been described to play an important role beyond the boundaries of the hospital setting, as indicated by the occurrence of community-associated infections in patients without discernible healthcare-associated risk factors (Coque et al, 2008; Doi et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Extended-Spectrum β-Lactamases (ESBL) and AmpC β-Lactamases in Enterobacterales: Development of a Screening Panel Using the MALDI-TOF MS-Based Direct-on-Target Microdroplet Growth Assay

et al. 2019

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Introduction: Antibiotic resistant bacteria are a growing concern worldwide. Extended-spectrum β-lactamases (ESBL) represent the most common resistance mechanism of Gram-negative bacteria against β-lactams, underlining the need for adequate diagnostic methods that provide reliable information in the shortest time possible. AmpC, a less prevalent but increasingly relevant class of β-lactamases, pose an additional challenge as their detection is complex. Here, we present an ESBL and AmpC screening panel employing the MALDI-TOF MS-based direct-on-target microdroplet growth assay (DOT-MGA).Materials and Methods: Four reference strains recommended by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) were used to develop the panel, which was further validated on 50 clinical Enterobacterales isolates resistant to third generation cephalosporins. The panel relies on the synergistic effect between ESBL and/or AmpC β-lactamase inhibitors and cephalosporins, which indicates β-lactamase production. Microdroplets containing the tested microorganism, cephalosporins in different concentrations and inhibitors were pipetted onto an MBT Biotarget and incubated for 3 or 4 h at 35 ± 1°C. Afterward, the liquid medium was removed and the material adhered to the spots was analyzed by MALDI-TOF MS. Synergy was detected by determining and comparing the minimum inhibitory concentrations of the tested cephalosporins with and without β-lactamase inhibitors. Data were interpreted following a diagnostic algorithm proposed by EUCAST in order to establish a final diagnosis. In comparison, PCR, broth microdilution (BMD) and combination disk tests (CDT) were performed.Results: Compared to the PCR results, the following positive and negative percent agreement values (PPA/NPA) were obtained for each resistance mechanism: ESBL, 94.44/100%; AmpC, 94.44/93.75% and ESBL+AmpC, 100/100%. These results, obtained after 4 h of incubation, were comparable with those of BMD and showed a higher accuracy than CDT.Discussion: We propose a novel phenotypic method for detection of ESBL and AmpC β-lactamases in Enterobacterales that provides reliable results in a short time, representing a promising alternative to the diagnostic techniques currently available. This easy-to-perform approach has potential for being implemented in routine laboratories, contributing to the further diversification of mass spectrometry technology into other fields such as antibiotic resistance testing.

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Detection of expanded-spectrum β-lactamases in Gram-negative bacteria in the 21st century

Cited by 20 publications

References 199 publications

Resistant Gram-Negative Bacteria and Diagnostic Point-of-Care Options for the Field Setting during Military Operations

Resistant Gram-Negative Bacteria and Diagnostic Point-of-Care Options for the Field Setting during Military Operations

More Pathogenicity or Just More Pathogens?—On the Interpretation Problem of Multiple Pathogen Detections with Diagnostic Multiplex Assays

Rapid Detection of Extended-Spectrum β-Lactamases (ESBL) and AmpC β-Lactamases in Enterobacterales: Development of a Screening Panel Using the MALDI-TOF MS-Based Direct-on-Target Microdroplet Growth Assay

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