Evaluation of Genotypic and Phenotypic Methods to Detect Carbapenemase Production in Gram-Negative Bacilli

McMullen, Allison R.; Yarbrough, Melanie L.; Wallace, Meghan A.; Shupe, Angela; Burnham, Carey Ann D.

doi:10.1373/clinchem.2016.264804

Cited by 29 publications

(25 citation statements)

References 30 publications

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“…We screened all samples with consensus primers targeting bla CTX − M , bla SHV , and bla TEM genes. All isolates were evaluated for carbapenemase production by using CARBA-NP test (bioMérieux, La Balme-les-Grottes, France) and GeneXpert system (Cepheid, Sunnyvale, USA) that targets bla KPC , bla NDM , bla IMP , bla VIM , and bla OXA-48-like genes [24,25]. The presence of carbapenemase-encoding genes (bla KPC , bla NDM , bla VIM , bla IMP , bla OXA-48-like , and bla IMI ) was further confirmed by PCR and sequencing for all isolates by the French Associated National Reference Centre for Antibiotic Resistance (FANRCAR, [26]).…”

Section: Esbl and Carbapenemase Identificationmentioning

confidence: 99%

Carbapenemase-producing Enterobacteriaceae circulating in the Reunion Island, a French territory in the Southwest Indian Ocean

Miltgen

Cholley

Martak

et al. 2020

Antimicrob Resist Infect Control

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Background: The spread of carbapenemase-producing Enterobacteriaceae (CPE) in the Southwest Indian Ocean area (SIOA) is poorly documented. Reunion Island is a French overseas territory located close to Madagascar and connected with Southern Africa, Indian sub-continent and Europe, with several weekly flights. Here we report the results of the CPE surveillance program in Reunion Island over a six-year period. Methods: All CPE were collected between January 2011 and December 2016. Demographics and clinical data of the carrier patients were collected. We determined their susceptibility to antimicrobials, identified the carbapenemases and ESBL by PCR and sequencing, and explored their genetic relationship using pulsed-field gel electrophoresis and multi-locus sequence typing. Results: A total of 61 CPEs isolated from 53 patients were retrieved in 6 public or private laboratories of the island. We found that 69.8% of CPE patients were linked to a foreign country of SIOA and that almost half of CPE cases (47.2%) reached the island through a medical evacuation. The annual number of CPE cases strongly increased over the studied period (one case in 2011 vs. 21 cases in 2016). A proportion of 17.5% of CPE isolates were nonsusceptible to colistin. bla NDM was the most frequent carbapenemase (79.4%), followed by bla IMI (11.1%), and bla IMP-10 (4.8%). Autochtonous CPE cases (30.2%) harboured CPE isolates belonging to a polyclonal population. Conclusions: Because the hospital of Reunion Island is the only reference healthcare setting of the SIOA, we can reasonably estimate that its CPE epidemiology reflects that of this area. Mauritius was the main provider of foreign CPE cases (35.5%). We also showed that autochthonous isolates of CPEs are mostly polyclonal, thus unrelated to cross-transmission. This demonstrates the local spread of carbapenemase-encoding genes (i.e. bla NDM) in a polyclonal bacterial population and raises fears that Reunion Island could contribute to the influx of NDMcarbapenemase producers into the French mainland territory.

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Section: Esbl and Carbapenemase Identificationmentioning

confidence: 99%

Carbapenemase-producing Enterobacteriaceae circulating in the Reunion Island, a French territory in the Southwest Indian Ocean

Miltgen

Cholley

Martak

et al. 2020

Antimicrob Resist Infect Control

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“…High-level temocillin resistance is a useful phenotypic trait for OXA-48 detection (21). Culture of rectal swab specimens followed by confirmation testing is a long process and often is not compatible with rapid implementation of reinforced hygiene measures (22).…”

Section: Nases) (3) the Ambler Class A Carbapenemases Include The Womentioning

confidence: 99%

“…Molecularly based techniques such as PCR and whole-genome sequencing remain the gold standard for the precise identification of carbapenemase genes (22,23). Molecular methods are now available for detecting carbapenemase genes from bacterial cultures but also directly from clinical specimens in less than an hour (22). Several assays are commercially available.…”

Section: Nases) (3) the Ambler Class A Carbapenemases Include The Womentioning

confidence: 99%

Evaluation of the Revogene Carba C Assay for Detection and Differentiation of Carbapenemase-Producing Gram-Negative Bacteria

et al. 2020

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The Revogene Carba C assay (formerly GenePOC Carba assay) is a multiplex nucleic acid-based in vitro diagnostic test intended for the detection of carbapenemase-producing Enterobacterales (CPE) from cultured colonies. This assay was evaluated directly on colonies of 118 well-characterized Enterobacterales with reduced susceptibility to carbapenems and on 49 multidrug-resistant (MDR) Pseudomonas aeruginosa and 40 MDR Acinetobacter baumannii isolates. The Revogene Carba C assay’s performance was high, as it was able to detect the five major carbapenemases (NDM, VIM, IMP, KPC, and OXA-48). In Enterobacterales, sensitivity and specificity were 100%. When extrapolating the results to the French CPE epidemiology between 2012 and 2018, this assay would have detected 99.28% of the 9,624 CPE isolates sent to the French NRC, missing 69 CPE isolates (2 GES-5, 10 OXA-23, 2 TMB-1, 1 SME-4, 53 IMI, and 1 FRI). The overall sensitivity and specificity for CP P. aeruginosa were 93.7 and 100%, respectively, as two rare IMP variants (IMP-31 and -46) were not detected. Extrapolating these results to the French epidemiology of CP P. aeruginosa in 2017, 93.3% would have been identified, missing only 1 DIM and 10 GES variants. The Revogene Carba C assay accurately identified the targeted carbapenemase genes in A. baumannii, but when extrapolating these results to the French CP A. baumannii epidemiology of 2017, only 12.50% of them could be detected, as OXA-23 is the most prevalent carbapenemase in CP A. baumannii. The Revogene Carba C assay showed excellent sensitivity and specificity for the five most common carbapenemases regardless of the bacterial host. It is well adapted to the CPE and CP P. aeruginosa epidemiology of many countries worldwide, which makes it suitable for use in the routine microbiology laboratory, with a time to result of ca. 85 min for eight isolates simultaneously.

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“…guidelines, the isolate is considered aztreonam susceptible but ceftazidime, meropenem, and imipenem resistant, a resistance profile suggestive of a Class B carbapenemase. The isolate was also positive for phenotypic production of a carbapenemase via the carbapenem inactivation method (4). To identify known carbapenemase genes in S. putrefaciens , we extracted genomic DNA with the Bacteremia kit (MoBIO, Carlsbad, CA), used 0.5 ng of DNA to make a Nextera XT Illumina sequencing library (5), and performed whole-genome sequencing on an Illumina NextSeq high-output sequencer to obtain 1,454,810 2 × 150 bp reads.…”

Section: Genome Announcementmentioning

confidence: 99%

Draft Genome Sequence of the bla _OXA-436 - and bla _NDM-1 -Harboring Shewanella putrefaciens SA70 Isolate

et al. 2017

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Evaluation of Genotypic and Phenotypic Methods to Detect Carbapenemase Production in Gram-Negative Bacilli

Abstract: Screening by CIM followed by the Xpert Carba-R PCR is an accurate method for detecting and characterizing CP-GNB, including ,, and complex.

Cited by 29 publications

References 30 publications

Carbapenemase-producing Enterobacteriaceae circulating in the Reunion Island, a French territory in the Southwest Indian Ocean

Carbapenemase-producing Enterobacteriaceae circulating in the Reunion Island, a French territory in the Southwest Indian Ocean

Evaluation of the Revogene Carba C Assay for Detection and Differentiation of Carbapenemase-Producing Gram-Negative Bacteria

Draft Genome Sequence of the bla _OXA-436 - and bla _NDM-1 -Harboring Shewanella putrefaciens SA70 Isolate

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Evaluation of Genotypic and Phenotypic Methods to Detect Carbapenemase Production in Gram-Negative Bacilli

Abstract: Screening by CIM followed by the Xpert Carba-R PCR is an accurate method for detecting and characterizing CP-GNB, including ,, and complex.

Cited by 29 publications

References 30 publications

Carbapenemase-producing Enterobacteriaceae circulating in the Reunion Island, a French territory in the Southwest Indian Ocean

Carbapenemase-producing Enterobacteriaceae circulating in the Reunion Island, a French territory in the Southwest Indian Ocean

Evaluation of the Revogene Carba C Assay for Detection and Differentiation of Carbapenemase-Producing Gram-Negative Bacteria

Draft Genome Sequence of the bla OXA-436 - and bla NDM-1 -Harboring Shewanella putrefaciens SA70 Isolate

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Draft Genome Sequence of the bla _OXA-436 - and bla _NDM-1 -Harboring Shewanella putrefaciens SA70 Isolate