The performance of a resazurin chromogenic agar plate with a combined disc method for rapid screening of extended‐spectrum‐β‐lactamases, AmpC β‐lactamases and co‐β‐lactamases in <i>Enterobacteriaceae</i>

Teethaisong, Yothin; Evans, Katie; Nakouti, Ismini; Tiamyom, Kanokwan; Cairns, James R. Ketudat; Hobbs, Glyn; Eumkeb, Griangsak

doi:10.1111/1348-0421.12499

Cited by 5 publications

(2 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A total of 111 well-documented 40,41 bacterial isolates from a well characterized collection of clinical isolates sourced in the UK between 2012-2017 were tested. The isolates included E. coli (n=37), K. pneumoniae (n=27), Enterobacter cloacae (n=15), Enterobacter aerogenes (n=12), Citrobacter freundii (n=12), Pseudomonas aeruginosa (n=3), Morganella morganii (n=3), K. oxytoca (n=1) and Acinetobacter haemolyticus (n=1).…”

Section: Methodsmentioning

confidence: 99%

A highly multiplexed melt-curve assay for detecting the most prevalent carbapenemase, ESBL and AmpC genes

Edwards

Williams

Teethaisong

et al. 2019

Preprint

View full text Add to dashboard Cite

AbstractResistance to third generation cephalosporins and carbapenems in Gram-negative bacteria is chiefly mediated by beta-lactamases including ESBL, AmpC and carbapenemase enzymes. Routine phenotypic detection methods do not provide timely results, and there is a lack of comprehensive molecular panels covering all important markers.An ESBL/carbapenemase HRM assay (SHV, TEM, CTX-M ESBLs, and NDM, IMP, KPC, VIM and OXA-48-like carbapenemases) and an AmpC HRM assay (16S rDNA control, FOX, MOX, ACC, EBC, CIT and DHA) were designed, and evaluated on 111 Gram-negative isolates with mixed resistance patterns.The sensitivity for carbapenemase, ESBL and AmpC genes was 96.7% (95%CI:82.8-99.9%), 93.6% (95%CI:85.7-97.9%) and 93.8% (95%CI:82.8-98.7%), respectively with a specificity of 100% (95%CI:95.6-100%), 93.9% (95%CI:79.8-99.3%) and 93.7% (95%CI:84.5-98.2%).The HRM assays enable the simultaneous detection of the fourteen most important ESBL, carbapenemase and AmpC genes and could be used as a molecular surveillance tool or to hasten detection of AMR for treatment management.

show abstract

Section: Methodsmentioning

confidence: 99%

A highly multiplexed melt-curve assay for detecting the most prevalent carbapenemase, ESBL and AmpC genes

Edwards

Williams

Teethaisong

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…A total of 111 bacterial isolates were obtained from a collection of clinical isolates sourced in the UK between 2012 and 2017 UK 35 , 36 . The isolates included E. coli (n = 37), K. pneumoniae (n = 27), Enterobacter aerogenes (n = 12), Enterobacter cloacae (n = 15), Citrobacter freundii (n = 12), P. aeruginosa (n = 3), Morganella morganii (n = 3), K. oxytoca (n = 1) and A. haemolyticus (n = 1).…”

Section: Methodsmentioning

confidence: 99%

Speciation of common Gram-negative pathogens using a highly multiplexed high resolution melt curve assay

Edwards

Sasaki

Williams

et al. 2018

Sci Rep

Self Cite

View full text Add to dashboard Cite

The identification of the bacterial species responsible for an infection remains an important step for the selection of antimicrobial therapy. Gram-negative bacteria are an important source of hospital and community acquired infections and frequently antimicrobial resistant. Speciation of bacteria is typically carried out by biochemical profiling of organisms isolated from clinical specimens, which is time consuming and delays the initiation of tailored treatment. Whilst molecular methods such as PCR have been used, they often struggle with the challenge of detecting and discriminating a wide range of targets. High resolution melt analysis is an end-point qPCR detection method that provides greater multiplexing capability than probe based methods. Here we report the design of a high resolution melt analysis assay for the identification of six common Gram-negative pathogens; Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Pseudomonas aeruginosa, Salmonella Sp, and Acinetobacter baumannii, and a generic Gram-negative specific 16S rRNA control. The assay was evaluated using a well characterised collection of 113 clinically isolated Gram-negative bacteria. The agreement between the HRM assay and the reference test of PCR and sequencing was 98.2% (Kappa 0.96); the overall sensitivity and specificity of the assay was 97.1% (95% CI: 90.1–99.7%) and 100% (95% CI: 91.78–100%) respectively.

show abstract

Microtiter plate with built-in oxygen sensors: a novel approach to investigate the dynamics of Pseudomonas aeruginosa growth suppression in the presence of divalent cations and antibiotics

2022

View full text Add to dashboard Cite

The depletion of dissolved oxygen in a defined synthetic medium can be measured in real time, using a micro-well plate format, associated with a fluorescent plate reader. This technology is appropriate for investigating the effect of antibiotics on cell kinetics because there is a direct correlation between the latter and the amount of dissolved oxygen in the medium of an assay. In this study, the metabolic activity of the opportunistic human pathogen Pseudomonas aeruginosa PA01 was investigated using the OxoPlate OP96U optical sensor technology. The response of P. aeruginosa to aminoglycoside antibiotics when Ca2+and Mg2+ ions are present in the Evans defined synthetic medium was measured. The results revealed that the effect of antibiotics on P. aeruginosa is influenced by the concentration of divalent cations present in the test medium, although the efficiency of Ca2+ in supressing antibiotic activity was found to be greater than that of Mg2+. By comparison to tobramycin, the effect of amikacin is largely inhibited by the Ca2+and Mg2+concentrations. The study results underscore that the reliability of the observation of growth inhibitors is enhanced by the oxygen consumption measurements. Thus, the OxoPlate OP96U system is proven to be an accurate method to test the effectiveness of antibiotic treatments against P. aeruginosa.

show abstract

The performance of a resazurin chromogenic agar plate with a combined disc method for rapid screening of extended‐spectrum‐β‐lactamases, AmpC β‐lactamases and co‐β‐lactamases in Enterobacteriaceae

Cited by 5 publications

References 33 publications

A highly multiplexed melt-curve assay for detecting the most prevalent carbapenemase, ESBL and AmpC genes

A highly multiplexed melt-curve assay for detecting the most prevalent carbapenemase, ESBL and AmpC genes

Speciation of common Gram-negative pathogens using a highly multiplexed high resolution melt curve assay

Microtiter plate with built-in oxygen sensors: a novel approach to investigate the dynamics of Pseudomonas aeruginosa growth suppression in the presence of divalent cations and antibiotics

Contact Info

Product

Resources

About