Multicenter Evaluation of the Xpert Carba-R Assay for Detection of Carbapenemase Genes in Gram-Negative Isolates

Traczewski, Maria M.; Carretto, Edoardo; Cantón, Rafael; Moore, Nicholas; Team, Carba-R Study

doi:10.1128/jcm.00272-18

Cited by 84 publications

(48 citation statements)

References 28 publications

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“…In addition, both assays showed a specificity of 100% (95% CI, 91.8% to 100%). Our results with the Xpert Carba-R assay are in line with previous studies, in which the recently added OXA-181 and OXA-232 gene sequences were also detected (16)(17)(18). Interestingly, the sensitivity of the Xpert Carba-R assay for the IMP variants included in this study was 44.4% (95% CI, 18.9% to 73.3%), with results for IMP-8, IMP-13, IMP-14, IMP-22, and IMP-50 variants being negative after repeated testing.…”

Section: Discussionsupporting

confidence: 92%

“…Interestingly, the sensitivity of the Xpert Carba-R assay for the IMP variants included in this study was 44.4% (95% CI, 18.9% to 73.3%), with results for IMP-8, IMP-13, IMP-14, IMP-22, and IMP-50 variants being negative after repeated testing. This is in line with the manufacturer´s specifications, which state that only IMP-1-related variants can be detected (16)(17)(18). The variants not detected by the Xpert Carba-R assay all had nucleotide identity with IMP-1 of less than 93% (see Table S4 in the supplemental material).…”

Section: Discussionsupporting

confidence: 83%

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Rapid and Easy Detection of Carbapenemases in Enterobacterales in the Routine Laboratory Using the New GenePOC Carba/Revogene Carba C Assay

2019

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The novel, real-time PCR-based GenePOC Carba assay on the microfluidic revogene platform (GenePOC, Québec, Canada; now Meridian Bioscience, Cincinnati, OH, USA) was recently designed for the detection of blaKPC, blaNDM, blaVIM, blaOXA-48-like, and blaIMP. The goals of this study were to evaluate the performance of this assay, to assess its suitability for the routine microbiology laboratory, and to compare it to the Xpert Carba-R assay for the detection of carbapenemase-producing Enterobacterales (CPE) strains. The Xpert Carba-R assay (Cepheid) and the GenePOC Carba assay were challenged with a collection of 176 clinical Enterobacterales isolates. The collection included 133 CPE strains producing a total of 139 carbapenemases, including VIM (n = 48), OXA-48-like (n = 40), NDM (n = 29), KPC (n = 13), and IMP (n = 9). Six isolates produced two different carbapenemases, and 43 carbapenemase-negative isolates were included as negative controls. The overall sensitivity for carbapenemase detection was 96.4% (95% confidence interval [CI], 91.9% to 98.5%) for the Xpert Carba-R assay and 100% (95% CI, 97.3% to 100%) for the GenePOC assay. The four most common carbapenemases (NDM, KPC, OXA-48-like, and VIM) were detected with a sensitivity of 100% (95% CI, 97.1% to 100%) by the two tests, with all double carbapenemase producers being correctly detected by both assays. The sensitivity of the Xpert Carba-R assay for IMP was 44.4% (95% CI, 18.9% to 73.3%), while that of the GenePOC assay was 100% (95% CI, 70.1% to 100%). The specificity of both assays was 100% (95% CI, 91.8% to 100%). The GenePOC Carba assay showed excellent sensitivity and specificity for the five most common carbapenemases, including IMP variants. Its simplicity and short turnaround time make it suitable for use in the routine microbiology laboratory for CPE detection.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 83%

Rapid and Easy Detection of Carbapenemases in Enterobacterales in the Routine Laboratory Using the New GenePOC Carba/Revogene Carba C Assay

2019

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“…The universally recognized need for a reliable OXA-48 test is suggested by the number of different assays that have been proposed in the past few years. Nucleic acid-based testing includes an FDA-approved PCR that can identify KPC, NDM, VIM, IMP, and OXA-48 (Cepheid Xpert Carba-R assay), which has been shown to have 100% sensitivity and greater than 97.1% specificity (38). Recently a lateral flow assay was developed that can identify OXA-48 and OXA-163 family carbapenemases using anti-OXA-48 antibodies, along with a multiplex lateral flow assay that can detect OXA-48, NDM, and KPC carbapenemases and a third technique that can identify the five main carbapenemases: KPC, NDM, VIM, IMP-type, and OXA-48 (32,39,40).…”

Section: Discussionmentioning

confidence: 99%

Identification of the OXA-48 Carbapenemase Family by Use of Tryptic Peptides and Liquid Chromatography-Tandem Mass Spectrometry

et al. 2019

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Phenotypic detection of the OXA-48-type class D ␤-lactamases in Enterobacteriaceae is challenging. We describe a rapid (less than 90 min) assay for the identification of OXA-48 family carbapenemases in subcultured bacterial isolates based on a genoproteomic approach. Following in silico trypsin digestion to ascertain theoretical core peptides common to the OXA-48 family, liquid chromatography-tandem mass spectrometry (LC-MS/MS) data-dependent acquisition was used to identify candidate peptide markers. Two peptides were selected based on performance characteristics: ANQAFLPASTFK, a core peptide common to all 12 OXA-48 family ␤-lactamase members, and YSVVPVYQEFAR, a highly specific peptide common to 11 of 12 OXA-48 family proteins providing the basis for an LC-MS/MS multiple reaction monitoring assay. An accuracy assessment was performed that included 98 isolates, 26 of which were OXA-48 positive. Two additional specificity assessments were performed including a mixture of isolates positive for OXA-48, KPC, NDM, VIM, and IMP carbapenemases. A combination of expert rules and expert judgment was applied by blinded operators to identify positive isolates. All isolates containing an OXA-48 family carbapenemase across all three test sets were correctly identified with no false positives, demonstrating 100% sensitivity (95% confidence interval [CI], 91.2% to 100%) and 100% specificity (95% CI, 96.2% to 100%) for the assay. These findings provide a framework for an LC-MS/MS-based method for the direct detection of OXA-48 family carbapenemases from cultured isolates that may have utility in predicting carbapenem resistance and tracking hospital outbreaks of OXA-48-carrying organisms.

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“…The proposed method was applied in a random selection of 143 clinical isolates previous molecular characterized by using the Xpert Carba-R Assay (Cepheid, Sunnyvale, United States), carrying 97 bla KPC , 1 bla GES , 12bla VIM , 4bla IMP , 3bla NDM , and 26bla OXA−48−like (Traczewski et al, 2018). The isolates were collected during a 2 months period (December 2017-January 2018) from 31 hospitals from Spain.…”

Section: Bacterial Isolatesmentioning

confidence: 99%

Rapid Detection of KPC-Producing Enterobacterales Susceptible to Imipenem/Relebactam by Using the MALDI-TOF MS MBT STAR-Carba IVD Assay

2020

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KPC-producing Enterobacterales represent a serious public health concern. Limited therapeutic options are available for treatment, however, the novel combination of imipenem/relebactam represents a promising alternative. To preserve the activity of this new antibiotic combination, only targeted treatments will be recommended, and rapid tests to detect susceptible bacteria are therefore urgently needed. Here, we propose a MALDI-TOF-based method using the MBT STAR-Carba IVD assay, Bruker Daltonik, to detect KPC-producing Enterobacterales susceptible to imipenem/relebactam in a random selection of 143 clinical isolates previous molecular characterized, carrying 97 bla KPC , 1 bla GES , 12bla VIM , 4bla IMP , 3bla NDM , and 26bla OXA−48−like . Species identification was confirmed by MALDI-TOF MS. The molecular characterization of the isolates was performed by the Xpert Carba-R Assay and the results were used as gold standard. Besides, all isolates were submitted to imipenem and imipenem/relebactam microdilution susceptibility testing. The assay showed an overall sensitivity and specificity to detect class A-producing Enterobacterales susceptible to imipenem/relebactam of 98% (96/98) and 93% (42/45), respectively. This MALDI-TOF-based methodology, with a turnaround time of less than 1 h, is a reliable test for detecting imipenem/relebactam activity and its inclusion in routine laboratory screening would facilitate the correct use of this new combination of antimicrobials as a targeted treatment.

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Multicenter Evaluation of the Xpert Carba-R Assay for Detection of Carbapenemase Genes in Gram-Negative Isolates

Cited by 84 publications

References 28 publications

Rapid and Easy Detection of Carbapenemases in Enterobacterales in the Routine Laboratory Using the New GenePOC Carba/Revogene Carba C Assay

Rapid and Easy Detection of Carbapenemases in Enterobacterales in the Routine Laboratory Using the New GenePOC Carba/Revogene Carba C Assay

Identification of the OXA-48 Carbapenemase Family by Use of Tryptic Peptides and Liquid Chromatography-Tandem Mass Spectrometry

Rapid Detection of KPC-Producing Enterobacterales Susceptible to Imipenem/Relebactam by Using the MALDI-TOF MS MBT STAR-Carba IVD Assay

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