Fast and reliable detection of carbapenemase genes in various Gram negatives using a new commercially available fluorescence-based real-time polymerase chain reaction platform

Sadek, Mustafa; Demord, Anthony; Poirel, Laurent; Nordmann, Patrice

doi:10.1016/j.diagmicrobio.2020.115127

Cited by 6 publications

(4 citation statements)

References 12 publications

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“…If detection of carbapenemase strains can help prevent outbreaks of and treat A. baumannii infections, then rapid organism and carbapenemase detection method is recommended and should be further investigated. Current strategies that have been recently developed include bioluminescence-based carbapenem susceptibility detection assay (BCDA), an accurate phenotypic method that allows detection of carbapenem-resistant species in 2Á5 h from culture media (van Almsick et al 2018), multiplex real-time PCR to detect and distinguish the most common carbapenemases present in A. baumannii (OXA-23like, OXA-24-like and OXA-58-like subfamilies) in 70 min even when more than one is simultaneously present (Mentasti et al 2020), Revogeneâ Carba C assay which is another a real-time PCR-based assay for the detection of genes that encode carbapenemases such as NDM, VIM, IMP, KPC and OXA-48 like) from various Gram-negative pathogens with a run time of about 70 min (Sadek et al 2020); fluorescence identification of b-lactamase activity (FIBA) to detect carbapenemase production in bacteria in 10 min (Feng et al 2020) or automated BD Phoenix CPO detect test (BD-CPO test) for the simultaneous detection and Ambler classification of carbapenemases in Enterobacteriaceae, P. aeruginosa and A. baumannii complex (results are accessible in 18 h from cultured bacterial growth) (Saad Albichr et al…”

Section: Roommentioning

confidence: 99%

Carbapenem resistance in Acinetobacter baumannii , and their importance in hospital‐acquired infections: a scientific review

2021

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Carbapenem is an important therapy for serious hospital-acquired infections and for the care of patients affected by multidrug-resistant organisms, specifically Acinetobacter baumannii; however, with the global increase of carbapenem-resistant A. baumannii, this pathogen has significantly threatened public health. Thus, there is a pressing need to better understand this pathogen in order to develop novel treatments and control strategies for dealing with A. baumannii. In this review, we discuss an overview of carbapenem, including its discovery, development, classification and biological characteristics, and its importance in hospital medicine especially in critical care units. We also describe the peculiarity of bacterial pathogen, A. baumannii, including its commonly reported virulence factors, environmental persistence and carbapenem resistance mechanisms. In closing, we discuss various control strategies for overcoming carbapenem resistance in hospitals and for limiting outbreaks. With the appearance of strains that resist carbapenem, the aim of this review is to highlight the importance of understanding this increasingly problematic healthcare-associated pathogen that creates significant concern in the field of nosocomial infections and overall public health. Carbapenem Development and history of carbapenemSafe and effective antibiotics play an important role in medical care and their development remains one of the

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Section: Roommentioning

confidence: 99%

Carbapenem resistance in Acinetobacter baumannii , and their importance in hospital‐acquired infections: a scientific review

2021

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“…Biological manufacturers have developed several polymer-based microfluidic commercial devices [e.g., Revogene (Meridian Bioscience, Cincinnati, OH, USA) and GenPlex ® (BOHUI, Beijing, China)] for the diagnosis of infectious diseases [ 1 , 71 , 72 , 73 ]. Nevertheless, weakness of polymer materials includes thermal conductivity and low thermal resistance.…”

Section: Multiplex Nucleic Acid Sensors On Microfluidic Platformsmentioning

confidence: 99%

Multiplex Detection of Infectious Diseases on Microfluidic Platforms

Chen

et al. 2023

Biosensors

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Infectious diseases contribute significantly to the global disease burden. Sensitive and accurate screening methods are some of the most effective means of identifying sources of infection and controlling infectivity. Conventional detecting strategies such as quantitative polymerase chain reaction (qPCR), DNA sequencing, and mass spectrometry typically require bulky equipment and well-trained personnel. Therefore, mass screening of a large population using conventional strategies during pandemic periods often requires additional manpower, resources, and time, which cannot be guaranteed in resource-limited settings. Recently, emerging microfluidic technologies have shown the potential to replace conventional methods in performing point-of-care detection because they are automated, miniaturized, and integrated. By exploiting the spatial separation of detection sites, microfluidic platforms can enable the multiplex detection of infectious diseases to reduce the possibility of misdiagnosis and incomplete diagnosis of infectious diseases with similar symptoms. This review presents the recent advances in microfluidic platforms used for multiplex detection of infectious diseases, including microfluidic immunosensors and microfluidic nucleic acid sensors. As representative microfluidic platforms, lateral flow immunoassay (LFIA) platforms, polymer-based chips, paper-based devices, and droplet-based devices will be discussed in detail. In addition, the current challenges, commercialization, and prospects are proposed to promote the application of microfluidic platforms in infectious disease detection.

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“…Metalloenzymes belong to the molecular B class (MBL). Considering their amino acid identity, carbapenemases are part of classes A, B and D (2)(3)(4). Class A members hydrolyze carbapenems, cephalosporins, penicillins, aztreonam.…”

Section: Introductionmentioning

confidence: 99%

“…A new multiplex fluorescence-based RT-PCR which runs on microfluidic Revogene platform was developed and is designed for the detection of genes encoding five carbapenemases: NDM, VIM, IMP, KPC and OXA-48 from Enterobacterales, Pseudomonas aeruginosa and Acinetobacter baumannii. The investigators reported a 100% sensitivity and specificity and characterized this assay as suitable for microbiology laboratories (4). The objectives of our study were: i) to identify the carbapenemase type using a multiplex RT-PCR with fluorescent labeled hydrolysis probes; to our knowledge this technique is used for the first time in Romania; ii) to test agreement between the types of carbapenemase identified by phenotypic methods and the detected carbapenemase genes.…”

Section: Introductionmentioning

confidence: 99%

Carbapenem resistance determinants in Klebsiella pneumoniae strains isolated from blood cultures-comparative analysis of molecular and phenotypic methods

TOMPA

Iancu

Pandrea

et al. 2022

Revista Romana De Medicina De Laborator

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Introduction: This study provides data on carbapenemases identified in carbapenem-resistant Klebsiella pneumoniae (CR-KP) isolated from blood-cultures by the multiplex molecular method. Material and method: Between October 2016 and September 2017, 47 non-duplicate Klebsiella pneumoniae (KP) were isolated from blood cultures, from hospitalized patients in the Regional Institute of Gastroenterology and Hepathology, Cluj-Napoca, Romania. Identification and antimicrobial susceptibility tests (AST) were performed by Vitek 2 Compact. The combination disks test (CDT) was used for phenotypic analysis and the LightCycler® Multiplex DNA assay was used to detect and identify the carbapenemases by the LightCycler®z 480 Instrument. The following targets were chosen: blaKPC, blaNDM, blaGES, blaIMP and blaOXA-48 genes and the Cobas® 4800 software variant 2.2.0 was used for the results interpretation. Results: Taking into consideration the meropenem minimum inhibitory concentration (MIC), 29 KP were susceptible and 18 were not-susceptible (MIC≥0.5 µg ml-1). In the CR-KP group, the CDT identified OXA-48 (10/18) and KPC (7/18) producers. One isolate showed a noninterpretable profile. The multiplex molecular analyses confirmed the carbapenemases production as: 9 CR-KP were KPC and OXA-48 co-producers, 8 were OXA-48 and one was KPC producing strains. In CR-KP group, we found a significant correlation between the CDT and RT-PCR tests results, concerning KPC (p = 0.671). Eight phenotypic results were confirmed by molecular Light-Cycler® Multiplex DNA assay. For CR-KP co-producers (KPC and OXA-48), the CDT could indicate only one carbapenem-hydrolyzing enzyme. Conclusion: This study highlights the CR-KP co-producers (OXA-48 and KPC). OXA-48-like is more frequently encountered in our area than other carbapenemases.

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Fast and reliable detection of carbapenemase genes in various Gram negatives using a new commercially available fluorescence-based real-time polymerase chain reaction platform

Cited by 6 publications

References 12 publications

Carbapenem resistance in Acinetobacter baumannii , and their importance in hospital‐acquired infections: a scientific review

Carbapenem resistance in Acinetobacter baumannii , and their importance in hospital‐acquired infections: a scientific review

Multiplex Detection of Infectious Diseases on Microfluidic Platforms

Carbapenem resistance determinants in Klebsiella pneumoniae strains isolated from blood cultures-comparative analysis of molecular and phenotypic methods

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