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Carbapenemase

Carbapenemase is an enzyme produced by certain bacteria that makes them resistant to the carbapenem class of antibiotics, which are often used as a last resort for treating severe infections.
These enzymes can break down and inactivate carbapenems, rendering them ineffective.
Carbapenemase-producing bacteria pose a significant public health threat due to their ability to cause difficult-to-treat infections.
Understanding the mechanisms and prevalencce of carbapenemase production is crucial for developing effective antimicrobial stewardship strategies and preventing the spread of these resistant pathogens.

Most cited protocols related to «Carbapenemase»

1
Carbapenemase Inactivation Method (CIM)
Cited 27 times
To perform the CIM, a suspension was made by suspending a full 10 μl inoculation loop of culture, taken from a Mueller-Hinton or blood agar plate in 400 μl water. Subsequently, a susceptibility-testing disk containing 10 μg meropenem (Oxoid Ltd, Hampshire, United Kingdom) was immersed in the suspension and incubated for a minimum of two hours at 35°C. After incubation, the disk was removed from the suspension using an inoculation loop, placed on a Mueller-Hinton agar plate inoculated with a susceptible E. coli indicator strain (ATCC 29522) and subsequently incubated at 35°C. Inoculation of the Mueller-Hinton agar plate with the indicator strain was done with a suspension of OD595 1.25 (correlates with a McFarland value of 0.5) streaked in three directions using a sterile cotton swab. If the bacterial isolate produced carbapenemase, the meropenem in the susceptibility disk was inactivated allowing uninhibited growth of the susceptible indicator strain. Disks incubated in suspensions that do not contain carbapenemases yielded a clear inhibition zone. If results are required within the same day, they can be read after six hours, but within the setting of our laboratory, we prefer reading results after overnight incubation (Fig. 1).
van der Zwaluw K., de Haan A., Pluister G.N., Bootsma H.J., de Neeling A.J, & Schouls L.M. (2015). The Carbapenem Inactivation Method (CIM), a Simple and Low-Cost Alternative for the Carba NP Test to Assess Phenotypic Carbapenemase Activity in Gram-Negative Rods. PLoS ONE, 10(3), e0123690.
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Publication 2015
A-Loop Agar Bacteria Blood carbapenemase Escherichia coli Gossypium Meropenem Psychological Inhibition Sterility, Reproductive Strains Susceptibility, Disease Vaccination
2
Comprehensive Antimicrobial Resistance Profiling
Cited 21 times
When AMR detection is switched on, Kleborate screens for known acquired AMR determinants using a curated version of the CARD AMR nucleotide database (v3.0.8 downloaded February 2020; see doi.org/10.6084/m9.figshare.13256759.v1 for full details on curation). Genes are identified using nucleotide BLAST (and amino acid search with tBLASTx if no exact nucleotide match is found). Gene truncations and spurious hits are identified as described above for virulence genes. Unlike the acquired forms, the intrinsic variants of oqxAB, chromosomal fosA and ampH are not associated with clinical resistance in KpSC and are therefore not reported. However, SHV, LEN or OKP β-lactamase alleles intrinsic to KpSC species are known to confer clinical resistance to penicillins and are reported in the Bla_chr column. Acquired SHV variants, and individual SHV sequence mutations known to confer resistance to extended-spectrum β-lactams or β-lactamase inhibitors, are reported separately (see Supplementary Note 3, Supplementary Data 11 and 12 for details).
Chromosomally encoded mutations and gene loss or truncations known to be associated with AMR are reported for genomes identified as KpSC species. These include fluoroquinolone resistance mutations in GyrA (codons 83 and 87) and ParC (codons 80 and 84)76 (link), and colistin resistance from truncation or loss of MgrB and PmrB57 –59 (link) (defined as <90% amino acid sequence coverage). Mutations in the OmpK35 and OmpK36 osmoporins reportedly associated with reduced susceptibility to β-lactamases41 (link),42 (link) are also screened and reported for KpSC genomes, and include truncation or loss of these genes and OmpK36GD and OmpK36TD transmembrane β-strand loop insertions41 (link). SHV β-lactamase, GyrA, ParC and OmpK mutations are identified by alignment of the translated amino acid sequences against a reference using BioPython, followed by an interrogation of the alignment positions of interest (see Supplementary Note 3, Supplementary Data 11 and 12 for a list of relevant positions).
AMR genes and mutations are reported by drug class, with β-lactamases further categorized by enzyme activity (β-lactamase, ESBL or carbapenemase, with/without resistance to β-lactamase inhibitors). Horizontally acquired AMR genes are reported separately from mutational resistance and contribute to the AMR gene count; these plus chromosomal mutations count towards the number of acquired resistance classes (intrinsic SHV alleles, reported in Bla_chr column, are not included in either count). Resistance scores are calculated as follows: 0 = no ESBL or carbapenemase, 1 = ESBL without carbapenemase (regardless of colistin resistance); 2 = carbapenamase without colistin resistance (regardless of ESBL); 3 = carbapenemase with colistin resistance (regardless of ESBL).
Lam M.M., Wick R.R., Watts S.C., Cerdeira L.T., Wyres K.L, & Holt K.E. (2021). A genomic surveillance framework and genotyping tool for Klebsiella pneumoniae and its related species complex. Nature Communications, 12, 4188.
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Publication 2021
Alleles Amino Acids Amino Acid Sequence beta-Lactamase beta-Lactamase Inhibitors beta-Lactam Resistance carbapenemase Chromosomes Codon Colistin enzyme activity Fluoroquinolones Genes Genome inhibitors Lactams Mutation Nucleotides Penicillin Resistance Pharmaceutical Preparations Susceptibility, Disease Virulence
3
National Healthcare Safety Network Antimicrobial Resistance Surveillance
Cited 18 times
HAIs that occurred in 2011–2014 and were reported to the Device-Associated and Procedure-Associated Modules of the Patient Safety Component of NHSN6 –9 as of December 16, 2015, were included in this report. These HAIs were reported from acute care hospitals, LTACHs, and IRFs, and include central line–associated bloodstream infections (CLABSIs), catheter-associated urinary tract infections (CAUTIs), all surgical site infections (SSIs) following inpatient procedures with a primary closure technique, and ventilator-associated pneumonias (VAPs). VAP surveillance in adult locations was retired from NHSN in January 2013 and was replaced with the surveillance of ventilator-associated events (VAEs). Therefore, VAP data in this report are limited to events in 2011–2012, and this will be the last report to include such data. Postprocedure pneumonias, asymptomatic bacteremic urinary tract infections, and pediatric VAPs, each of which accounted for less than 1% of reported HAIs, were excluded from these analyses. NHSN surveillance methodology has been reported elsewhere6 –9 and is summarized in the first NHSN antimicrobial resistance report.1 (link)Pathogen and antimicrobial susceptibility data reported to NHSN are provided by the facility’s designated clinical microbiology laboratory. At most, 3 pathogens can be reported per HAI. For some pathogens, there is a select group of antimicrobials for which susceptibility test results must be reported if testing was performed. Laboratories are expected to use the current Clinical and Laboratory Standards Institute standards for antimicrobial susceptibility testing.10 Susceptibility results were reported using the category interpretations of susceptible [S], intermediate [I], resistant [R], or not tested. Because laboratories may test different antimicrobial agents within a class, for some phenotypes, resistance was defined using at least 1 of several agents within the same class.
Resistance for Staphylococcus aureus and Enterococcus spp. phenotypes included those pathogens that tested R to oxacillin, methicillin, or cefoxitin (methicillin-resistant S. aureus), or vancomycin (vancomycin-resistant Enterococcus). To be defined as resistant to extended-spectrum cephalosporins, pathogens must have tested I or R to either ceftazidime or cefepime (Pseudomonas aeruginosa) or to ceftazidime, cefepime, ceftriaxone, or cefotaxime (Enterobacteriaceae). Carbapenem resistance, as defined in this report, included all applicable pathogens with a result of I or R to imipenem, meropenem, or doripenem unless otherwise noted. Fluoroquinolone resistance was defined as a result of I or R to either ciprofloxacin or levofloxacin (P. aeruginosa) or to ciprofloxacin, levofloxacin, or moxifloxacin (Escherichia coli). Aminoglycoside resistance in P. aeruginosa was defined as a result of I or R to gentamicin, amikacin, or tobramycin. Finally, definitions of multidrug-resistance required a test result of I or R to at least 1 agent within a class—thus establishing nonsusceptibility to the class—and nonsusceptibility to at least 3 of the specified classes. For Enterobacteriaceae species and P. aeruginosa, 5 classes were included in the criteria: extended-spectrum cephalosporins, fluoroquinolones, aminoglycosides, carbapenems, and piperacillin or piperacillin/tazobactam. A sixth class, ampicillin/sulbactam, was included in the criteria for multidrug-resistance for Acinetobacter spp. These criteria approximated interim standard definitions for defining multidrug-resistance.11 (link) Results from Klebsiella pathogens were limited to K. pneumoniae and K. oxytoca combined; other species of Klebsiella were extremely rare and excluded from the analysis.
As discussed above, carbapenem-resistant Enterobacteriaceae (CRE) was defined in this report as any K. pneumoniae, K. oxytoca, E. coli, or Enterobacter spp. that tested I or R to imipenem, meropenem, or doripenem. However, this definition was updated in NHSN in 2015 to increase detection of carbapenemase-producing strains.12 (link)–14 To anticipate the impact of the updated CRE definition, the resistance percentages for CRE using both the current and updated definitions were calculated using 2014 data. In subsequent reports, CDC will use only the updated definition, which includes the above mentioned Enterobacteriaceae pathogens that test R to imipenem, meropenem, doripenem, or ertapenem.
Data were analyzed with SAS software, version 9.3 (SAS Institute). For reporting hospitals and all reported HAIs and pathogens, absolute frequencies and distributions were calculated by hospital characteristic, HAI, surgery, and location type. The 15 most frequently reported pathogens were identified, and their frequencies and ranks within each HAI or surgery type were calculated. For each HAI type and period, a pooled mean percent resistance was calculated for each pathogen-antimicrobial combination (ie, sum of pathogens that tested resistant, divided by the sum of pathogens tested for susceptibility, multiplied by 100). The pooled mean percent resistance was not calculated for any phenotype for which less than 20 pathogens were tested. In addition, the percentage of pathogens that were tested for susceptibility (sum of pathogens tested for susceptibility, divided by the sum of total pathogens isolated, multiplied by 100) was calculated for each pathogen–antimicrobial agent combination.
Statistical analyses were not performed to test for temporal changes in the resistance percentage in 2011–2014, and thus, this report does not convey any definitive conclusions regarding changes in resistance over time. The results and discussions presented in this paper are based solely on observed differences in the magnitude of the resistance percentage.
Weiner L.M., Webb A.K., Limbago B., Dudeck M.A., Patel J., Kallen A.J., Edwards J.R, & Sievert D.M. (2016). Antimicrobial-Resistant Pathogens Associated With Healthcare-Associated Infections: Summary of Data Reported to the National Healthcare Safety Network at the Centers for Disease Control and Prevention, 2011–2014. Infection control and hospital epidemiology, 37(11), 1288-1301.
Publication 2016
4
Validation of Carbapenemase Identification Method
Cited 17 times
For validation of the CIM, a selection of 30 Gram-negative isolates was used. This selection included isolates obtained from different institutes across the world carrying known carbapenemase encoding genes and carbapenem susceptible isolates, according to the submitter (Table 1). In addition, 694 isolates submitted to the National Institute for Public Health and the Environment for the national surveillance of carbapenemase-producing Enterobacteriaceae (CPE) by Dutch medical microbiology laboratories (MMLs) during the first six months of 2012 and the first six months of 2013 were used. For the national surveillance of CPE in the Netherlands, Dutch MMLs are requested to submit Enterobacteriaceae isolates with an MIC for meropenem > 0.25 μg/ml. However, more than half of the isolates (411/694, 59%) sent in for CPE surveillance were non-fermenting Gram-negatives belonging to the genera Pseudomonas and Acinetobacter. Furthermore, 35% of the isolates had MICs below 0.25 μg/ml. Nevertheless, all isolates were included in this study.
The species identification, as performed by the MMLs, was confirmed using MALDI-TOF (Bruker Daltonics GmbH, Bremen, Germany) and the MIC for all isolates was confirmed by E-test (BioMerieux Inc., Marcy L’Etoile, France). Culturing of isolates was done on Columbia Sheep Blood (bioTRADING Benelux BV, Mijdrecht, The Netherlands) and Mueller-Hinton agarplates (Oxoid Ltd, Hampshire, United Kingdom). An overview of all CPE surveillance isolates and their characteristics is displayed in Tables 2 and 3.
van der Zwaluw K., de Haan A., Pluister G.N., Bootsma H.J., de Neeling A.J, & Schouls L.M. (2015). The Carbapenem Inactivation Method (CIM), a Simple and Low-Cost Alternative for the Carba NP Test to Assess Phenotypic Carbapenemase Activity in Gram-Negative Rods. PLoS ONE, 10(3), e0123690.
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Publication 2015
Acinetobacter Blood Carbapenem-Resistant Enterobacteriaceae carbapenemase Carbapenems Domestic Sheep Enterobacteriaceae Genes Meropenem Pseudomonas Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
5
Insect Model for Pathogenic Bacteria
Cited 15 times
Each KPC(+) bloodstream isolate from the clinical study was randomly matched to one KPC(−) isolate from the clinical study. Twelve randomly selected insects weighing between 250–350 mg were selected for each isolate (Vanderhorst, Inc, St. Mary’s, OH). The insects were inoculated by injecting 1 x 106 CFU per 10 microliter aliquot into the hemoceol via the rear left proleg using a 10 μL Hamilton animal syringe (Hamilton Co., Reno, NV, USA). Colony counts were performed by serial dilution with ultimate plating on blood agar (Remel, Lenexa, KS). Individual colonies were enumerated after 18–24 hours incubation at 38°C in ambient air. Any experiment with a colony count outside of a half log10 deviation was repeated. Phosphate buffer solution (placebo) injection controls and controls receiving no injection were used to evaluate trauma and attrition, respectively. Results were not included if greater than or equal to two insects died in either of the control groups [15 (link)]. The insects were incubated at 37°C in atmospheric air and observed every 24 h for 5 days. One common isolate was used for each experiment to control for inter-experiment insect robustness. Experiments were performed in duplicate and repeated in the case of discordance. Representative results are reported for all final experiments meeting inclusion criteria.
McLaughlin M.M., Advincula M.R., Malczynski M., Barajas G., Qi C, & Scheetz M.H. (2014). Quantifying the clinical virulence of Klebsiella pneumoniae producing carbapenemase Klebsiella pneumoniae with a Galleria mellonella model and a pilot study to translate to patient outcomes. BMC Infectious Diseases, 14, 31.
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Publication 2014
Agar Animals BLOOD Blood Circulation Buffers Insecta Insect Control Phosphates Placebos Syringes Technique, Dilution Tooth Attrition Wounds and Injuries

Most recents protocols related to «Carbapenemase»

1
Carbapenemase Detection Protocol
A phenotypic confirmation of carbapenemase production was performed by the modified Hodge test and by the KPC&MBL&OXA-48 disk kit (Liofilchem, Roseto degli Abruzzi, Italy), acc. EUCAST, 2023.
Markovska R., Stankova P., Stoeva T., Keuleyan E., Mihova K, & Boyanova L. (2024). In Vitro Antimicrobial Activity of Five Newly Approved Antibiotics against Carbapenemase-Producing Enterobacteria—A Pilot Study in Bulgaria. Antibiotics, 13(1), 81.
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Publication 2024
2
Carbapenemase Detection by PCR and Immunoassay
Transconjugants were analyzed by PCR using Platinum™ Taq DNA polymerase (Invitrogen, ThermoFischer Scientific, USA) and specific primers for blaKPC−2 and blaVIM−1 (Table S3), as previously described [31 (link)]. Carbapenemase expression was confirmed by immunochromatography using NG-Test Carba 5 assay, according to the manufacturer’s instructions [32 (link)].
Gálvez-Silva M., Arros P., Berríos-Pastén C., Villamil A., Rodas P.I., Araya I., Iglesias R., Araya P., Hormazábal J.C., Bohle C., Chen Y., Gan Y.H., Chávez F.P., Lagos R, & Marcoleta A.E. (2024). Carbapenem-resistant hypervirulent ST23 Klebsiella pneumoniae with a highly transmissible dual-carbapenemase plasmid in Chile. Biological Research, 57, 7.
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Publication 2024
3
Screening for Carbapenemase Production
The phenotypic screening of carbapenemase production was done with the Triton Hodge Test and the carbapenem inactivation method as previously described (38 (link)). Inhibition tests using EDTA and phenylboronic acid were used to distinguish classes of carbapenemases (39 (link)).
Cuicapuza D., Loyola S., Velásquez J., Fernández N., Llanos C., Ruiz J., Tsukayama P, & Tamariz J. (2024). Molecular characterization of carbapenemase-producing Enterobacterales in a tertiary hospital in Lima, Peru. Microbiology Spectrum, 12(2), e02503-23.
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Publication 2024
4
Phenotypic Detection of Carbapenemase Production
Not available on PMC !
Modified hodge test (Mht) for phenotypic detection of carbapenemase production:
All carbapenem-resistant isolates were subjected to the MHT. A lawn culture of a 1:10 dilution of Escherichia coli ATCC 25922 was prepared on a Muller-Hinton agar plate, and a 10 µg meropenem disk was placed at the center of the plate. A straight line was drawn from the edge of the disk to the edge of the plate using the test organism. Four strains were tested on the same plate with one disk, and the plate was then incubated overnight at 35±2°C in an aerobic atmosphere. After 16-24 hours of incubation, the test result is interpreted as positive if there is a clover leaf-like indentation of the Escherichia coli ATCC 25922 strain growing along the test organism's streak within the disk diffusion zone, indicating the production of carbapenemase. A negative result indicates no growth of Escherichia coli ATCC 25922 along the test organism's streak within the disk diffusion zone [8, 9] (link). [Table /Fig 1] shows the MHT.
[Table /Fig All the carbapenem-resistant isolates were subjected to the double disk synergy test using Imipenem and EDTA. To prepare a 0.5
McFarland standard of the test isolate, 2 to 3 colonies of the test isolate were inoculated into peptone water and incubated for 2-3 hours at 37°C. After incubation, a lawn culture of the organism was inoculated onto MHA following the CLSI guidelines. A sterile cotton swab was then dipped into the 0.5 McFarland standard inoculum and streaked across the entire MHA plate. After drying, a 10 µg imipenem disc was placed on the lawn culture, maintaining a distance of 24 mm center to center from the imipenem-EDTA (10-750 µg) disc. The plate was then incubated at 35±2°C for 16 to 18 hours. The zone diameter was measured using a calibrated zone scale. A MBL positive strain was considered when the increase in the inhibition zone with the imipenem-EDTA disk was ≥7 mm compared to the imipenem disk alone [10] . [Table/ Fig 2] shows the DDT.
[ The shows that out of all clinical samples, 842 (66.5%) were Gram-negative isolates, and 424 (33.5%) were Gram-positive isolates.
Pilli R., Rani A.D., Sunitha B., & Venna S.C. (2024). Detection of Carbapenemase Producing Gram-negative Bacteria at a Tertiary Care Hospital: A Cross-sectional Study. National Journal of Laboratory Medicine.
Publication 2024
5
Phenotypic and Genotypic Detection of KPC Carbapenemase
Phenotypic detection of KPC carbapenemase was performed using the KPC/MBL/OXA-48 Confirm Kit (Rosco Diagnostica, Taastrup, Denmark) and the immunochromatography test O.K.N.V.I. RESIST-5 (CORIS BioConcept, Gembloux, Belgium). Susceptibility testing by double-disk synergy test (DDST) and ESBL/AmpC Screen Kit tests (Rosco Diagnostica) were also performed. KPC carbapenemase genes were detected by the Eazyplex-Superbug-CRE system (Amplex-Biosystems, Germany) and the Xpert Carba-R assay (Cepheid, Sunnyvale, USA) and then confirmed by PCR and Sanger sequencing.
Castillo-Polo J.A., Hernández-García M., Maruri-Aransolo A., de la Vega C., Ruiz-Garbajosa P, & Cantón R. (2024). Evolution of ceftazidime-avibactam and cefiderocol resistance in ST131-H30R1-Escherichia coli isolates with KPC-3 mutants and application of FTIR biotyping. Microbiology Spectrum, 12(4), e02776-23.
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Publication 2024

Top products related to «Carbapenemase»

1
Vitek 2 system by bioMérieux
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The Vitek 2 system is an automated microbiology platform designed for the rapid identification and antimicrobial susceptibility testing of microorganisms. The system utilizes miniaturized biochemical testing to provide accurate results for a wide range of bacterial and yeast species.
2
Etest by bioMérieux
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Etest is a quantitative antimicrobial susceptibility testing (AST) method developed by bioMérieux. It provides minimum inhibitory concentration (MIC) values for specific antimicrobial agents. Etest utilizes a predefined antimicrobial gradient on a plastic strip to determine the MIC of a tested microorganism.
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MALDI-TOF MS is a type of mass spectrometry instrument that uses Matrix-Assisted Laser Desorption/Ionization (MALDI) as the ionization technique and Time-of-Flight (TOF) as the mass analyzer. It is designed to analyze and identify a wide range of compounds, including proteins, peptides, lipids, and small molecules.
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More about "Carbapenemase"

Carbapenemase is a critical enzyme that poses a significant threat to public health due to its ability to render carbapenems, a class of potent antibiotics, ineffective against certain bacteria.
These enzymes, produced by various bacterial species, can break down and inactivate carbapenems, making it extremely challenging to treat severe infections caused by these resistant pathogens.
Understanding the mechanisms and prevalence of carbapenemase production is crucial for developing effective antimicrobial stewardship strategies and preventing the spread of these resistant microbes.
Diagnostic tools like the Vitek 2 system, Etest, MALDI-TOF MS, and the VITEK 2 Compact system play a vital role in identifying and characterizing carbapenemase-producing bacteria.
The QIAamp DNA Mini Kit is often used for efficient DNA extraction from these resistant strains, enabling further genetic analysis and characterization.
The VITEK MS, a MALDI-TOF mass spectrometry system, can also be employed to rapidly identify and differentiate carbapenemase-producing organisms.
Additionally, the Vitek 2 automated system is a powerful tool for susceptibility testing, helping to determine the most effective antibiotics, including meropenem, for treating infections caused by carbapenemase-producing bacteria.
The MALDI Biotyper is another advanced technology that can be utilized to accurately identify and distinguish carbapenemase-producing isolates, aiding in the development of targeted treatment plans and infection control measures.
By leveraging these state-of-the-art diagnostic tools and techniques, researchers and clinicians can gain crucial insights into the mechanisms, prevalence, and spread of carbapenemase-producing bacteria, ultimately paving the way for more effective strategies to combat this growing public health challenge.