Direct-PCR from rectal swabs and environmental reservoirs: A fast and efficient alternative to detect blaOXA-48 carbapenemase genes in an Enterobacter cloacae outbreak setting

Probst, Katja; Boutin, Sébastien; Späth, Isabel; Scherrer, M; Henny, Nicole; Sahin, Delal; Heininger, Alexandra; Heeg, Klaus; Nurjadi, Dennis

doi:10.1016/j.envres.2021.111808

Cited by 6 publications

(4 citation statements)

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“…Antibiotic susceptibility testing was performed on the Vitek 2 (bioMérieux) and interpreted according to the EUCAST clinical breakpoints of the respective year of detection. The presence of carbapenemase was detected using an in-house PCR, as described elsewhere ( 37 , 38 ).…”

Section: Methodsmentioning

confidence: 99%

“…Wastewater from toilets and shower drains was collected at defined intervals, weekly during the peak of the outbreak and otherwise monthly, from all patient rooms of the three affected wards, as described elsewhere ( 38 ). The total sampling volume was approximately 40 ml, and 10 μl of the samples was cultured using chromID Carba Smart agar plates (bioMérieux GmbH) and incubated at 37°C overnight.…”

Section: Methodsmentioning

confidence: 99%

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Genomic Investigation and Successful Containment of an Intermittent Common Source Outbreak of OXA-48-Producing Enterobacter cloacae Related to Hospital Shower Drains

et al. 2021

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Genomic Investigation and Successful Containment of an Intermittent Common Source Outbreak of OXA-48-Producing Enterobacter cloacae Related to Hospital Shower Drains

et al. 2021

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“…For instance, mPCR offers the advantage of the simultaneous detection of multiple resistant genes through the use of multiple sets of primers [ 99 , 118 , 123 , 131 ]. Real-time PCR, or qPCR, allows for the rapid simultaneous detection and quantification of amplified PCR products using fluorescent dyes, eliminating gel electrophoresis [ 99 , 132 , 133 ]. Automated systems of PCR or qPCR are commercially available and automatically purify the sample, concentrate DNA, and amplify and detect major bacterial genes, confirming antibiotic resistance in less than two hours [ 99 , 134 ].…”

Section: Current and Emerging Detection Techniques Of Crementioning

confidence: 99%

A Review of Carbapenem Resistance in Enterobacterales and Its Detection Techniques

Caliskan-Aydogan

Alocilja

2023

Microorganisms

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Infectious disease outbreaks have caused thousands of deaths and hospitalizations, along with severe negative global economic impacts. Among these, infections caused by antimicrobial-resistant microorganisms are a major growing concern. The misuse and overuse of antimicrobials have resulted in the emergence of antimicrobial resistance (AMR) worldwide. Carbapenem-resistant Enterobacterales (CRE) are among the bacteria that need urgent attention globally. The emergence and spread of carbapenem-resistant bacteria are mainly due to the rapid dissemination of genes that encode carbapenemases through horizontal gene transfer (HGT). The rapid dissemination enables the development of host colonization and infection cases in humans who do not use the antibiotic (carbapenem) or those who are not hospitalized but interacting with environments and hosts colonized with carbapenemase-producing (CP) bacteria. There are continuing efforts to characterize and differentiate carbapenem-resistant bacteria from susceptible bacteria to allow for the appropriate diagnosis, treatment, prevention, and control of infections. This review presents an overview of the factors that cause the emergence of AMR, particularly CRE, where they have been reported, and then, it outlines carbapenemases and how they are disseminated through humans, the environment, and food systems. Then, current and emerging techniques for the detection and surveillance of AMR, primarily CRE, and gaps in detection technologies are presented. This review can assist in developing prevention and control measures to minimize the spread of carbapenem resistance in the human ecosystem, including hospitals, food supply chains, and water treatment facilities. Furthermore, the development of rapid and affordable detection techniques is helpful in controlling the negative impact of infections caused by AMR/CRE. Since delays in diagnostics and appropriate antibiotic treatment for such infections lead to increased mortality rates and hospital costs, it is, therefore, imperative that rapid tests be a priority.

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“…The developed PCR method using primers bla SHV , bla CTX-M , bla TEM , and bla OXA and facilitated their simultaneous detection . Several instances of carbapenemase-encoding genes detection using direct methods have been reported(Nijhuis 2013; Pery 2018;Bordin 2019;Cury 2020;Probst 2022) . Nijhuis et al used bla KPC , bla OXA-48 , bla VIM , and bla NDM primers to detect carbapenemase-producing isolates by real-time PCR using the commercial product, Check-Direct CPE assay (Check-Points, Wageningen, Netherlands) .…”

mentioning

confidence: 99%

Simple and quick detection of extended-spectrum <i>β</i>-lactamase and carbapenemase-encoding genes using isothermal nucleic acid amplification techniques

NAKAYAMA,

SOGA

2023

Journal of Microorganism Control

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The spread of plasmid-mediated antibiotic-resistant bacteria must be controlled; to this end, developing kits for simple and rapid detection in food and clinical settings is desirable. This review describes the detection of antibiotic resistance genes in extended-spectrum β-lactamase (ESBL) -and carbapenemase-producing bacteria. Loop-mediated isothermal amplification (LAMP) , a technique developed in Japan, is a useful diffusion amplification method that does not require equipment like thermal cyclers, and amplifies the target gene in 30 min at about 65℃. Although most reports targeting ESBL and carbapenemase genes are intended for clinical use, environmental and food samples have also been targeted. Recombinase polymerase amplification (RPA) has recently been developed; in RPA, the reaction proceeds under the human skin with reaction conditions of 30 min at 37℃. Detection of ESBL and carbapenemase-encoding genes in food and clinical samples using RPA has been reported in limited studies. However, research on RPA has just begun, and further development is expected.

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Direct-PCR from rectal swabs and environmental reservoirs: A fast and efficient alternative to detect blaOXA-48 carbapenemase genes in an Enterobacter cloacae outbreak setting

Cited by 6 publications

References 41 publications

Genomic Investigation and Successful Containment of an Intermittent Common Source Outbreak of OXA-48-Producing Enterobacter cloacae Related to Hospital Shower Drains

Genomic Investigation and Successful Containment of an Intermittent Common Source Outbreak of OXA-48-Producing Enterobacter cloacae Related to Hospital Shower Drains

A Review of Carbapenem Resistance in Enterobacterales and Its Detection Techniques

Simple and quick detection of extended-spectrum <i>β</i>-lactamase and carbapenemase-encoding genes using isothermal nucleic acid amplification techniques

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