Types Of Carbapenemases Research Articles

Elucidating the resistance mechanisms and binding pattern of novel Oxa-48-like carbapenemases covalent inhibitors: A hybrid experimental and in silico approach

Klebsiella pneumoniae is a Gram negative bacteria that can cause harm to the public's health and vulnerability has increased due to the emergence of highly virulent strains resistant to carbapenem antibiotics. The K. pneumoniae resistance mechanism involves extended-spectrum beta-lactamases, which are enzymes that can break down a broad range of antibiotics. The particular concern is Class D β-lactamases like OXA-48, which further complicates treatment options for infections caused by K. pneumoniae. The current study screened 7800 samples from three hospitals in Pakistan and found 353 positive samples of K. pneumoniae. The isolates were resistant to SXT, AMP, FEP CTX, CIP, TET, MEM, and IPM, while effective antibiotics were TGC, FOS, FOX, SCF and TZP. The sequencing and mutational analysis of the OXA-48 gene revealed 45-point mutations, including five novel missense mutations (Thr104:Ala104, Asn110:Asp110, Glu168:Gln168, Ser171:Ala171 and Arg214:Ser214). These mutations can contribute to the antibiotic affinity decrease and cause resistance. Furthermore, the study aimed to develop covalent inhibitors of OXA-48 (wild type and mutant) carbapenemases using a combination of molecular docking and molecular dynamic (MD) simulation approaches from natural products from the ZNINC20 database. 10 compounds (ZINC103533306, ZINC103531564, ZINC3861001, ZINC103533375, ZINC12411532, ZINC103533290, ZINC12376465, ZINC85875819, ZINC59586513 and ZINC13370550) showed best fit and high interactions with the OXA-48 active pocket. The binding free energies calculated from the MD showed that these compounds have high affinity with both the wild type and mutated OXA-48 protein. The selected compounds showed good drug-like properties and can be the leading inhibitors to combat the resistant K. pneumoniae strains.

Carbapenem resistance in Enterobacterales: Predicting clinical outcomes in bloodstream infections

PurposeCarbapenem-resistant Enterobacterales (CRE) are a global concern due to their high mortality rates and limited therapeutics. CRE-caused bloodstream infections (BSIs) are challenging to manage, especially in healthcare settings. This study aimed to investigate the predictors of mortality in BSI patients caused by CRE. MethodsA single center prospective study to examine the characteristics of BSI caused by CRE in a large academic hospital over 15 months. The main outcomes were microbiological characteristics and clinical outcomes of patients at 28 days based on a step-wise regression analysis. ResultsA total of 76 episodes of BSI due to CRE were included. The study found that the most common type of carbapenemase was OXA-48 (69.7 %, n = 53), followed by the co-existence of OXA-48 and MBL (26.3 %, n = 20), with Klebsiella pneumoniae being the most common (90 %, n = 69). Patients with OXA-48-BSI were more likely to have a urinary source of infection, while patients with MBL-BSI were more likely to have a non-urinary source of infection. All cases (100 %) had medical devices. Around 30.3 % of patients received effective empirical treatment, while 61.8 % received adequate therapy at 48 h. The overall mortality rate was 42.1 % (n = 32), and the main predictors of mortality in this study were the presence of sepsis and inadequate initial therapy, while age >65 predicted mortality in the linear regression but not the stepwise regression model. ConclusionCRE-BSIs are a serious health threat. The study highlights the need for preventive strategies focused on high-risk patients and proper device management to reduce BSI.

What do We Know so Far about Ges Carbapenemases, and What Threat do They Pose?

Abstract Carbapenemases, classified as bacterial enzymes, have the ability to hydrolyze carbapenems – important broad-spectrum antibiotics. This work attempts to summarize the information on the diversity of Guiana Extended-Spectrum (GES) subgroup of carbapenemases, and highlights the serious threat posed by infections caused by bacteria capable of producing these enzymes. The structure, functional characteristics, classification of different types of GES carbapenemases and diagnostic methods are discussed in detail. There are 59 GES-type carbapenemases, which have different amino acid sequences of the protein chains as well as activity against various antibiotics. Currently, bacterial strains with antibiotic resistance of the GES type are treated with: cefiderocil belonging to the cephalosporins, eravacycline belonging to the tetracyclines, lefamulin belonging to the pleuromutulins, colistin, fosfomycin, nitrofurantoin, tobramycin, amikacin, imipenem with relebactam, meropenem with waborbactam, ceftazidime with avibactam and plazomycin. In addition, the following drugs are under study: durlobactam with sulbactam, taniborbactam and cefepime with enmetazobactam This paper aims to summarize the current knowledge on GES-type carbapenemases, their diagnosis and treatment.

In vitro Synergistic and Bactericidal Effects of Aztreonam in Combination with Ceftazidime/ Avibactam, Meropenem/Vaborbactam and Imipenem/Relebactam Against Dual-Carbapenemase-Producing Enterobacterales.

Our aim was to elucidate the resistance mechanisms and assess the combined synergistic and bactericidal activities of aztreonam in combination with ceftazidime/avibactam (CZA), meropenem/vaborbactam (MEV), and imipenem/relebactam (IMR) against Enterobacterales strains producing dual carbapenemases. Species identification, antimicrobial susceptibility testing and determination of carbapenemase type were performed for these strains. Plasmid sizes, plasmid conjugation abilities and the localization of carbapenemase genes were investigated. Whole-genome sequencing was performed for all strains and their molecular characteristics were analyzed. In vitro synergistic and bactericidal activities of the combination of aztreonam with CZA, MEV and IMR against these strains were determined using checkerboard assay and time-kill curve assay. A total of 12 Enterobacterales strains producing dual-carbapenemases were collected, including nine K. pneumoniae, two P. rettgeri, and one E. hormaechei. The most common dual-carbapenemase gene pattern observed was bla (KPC-2+NDM-5) (n=4), followed by bla KPC-2+IMP-26 (n=3), bla (KPC-2+NDM-1) (n=2), bla (KPC-2+IMP-4) (n=1), bla (NDM-1+IMP-4) (n=1) and bla (KPC-2+KPC-2) (n=1). In each strain, the carbapenemase genes were found to be located on two distinct plasmids which were capable of conjugating from the original strain to the receipt strain E. coli J53. The results of the checkerboard synergy analysis consistently revealed good synergistic effects of the combination of ATM with CZA, MEV and IMR. Except for one strain, all strains exhibited significant synergistic activity and bactericidal activity between 2 and 8 hours. Dual-carbapenemase-producing Enterobacterales posed a significant threat to clinical anti-infection treatment. However, the combination of ATM with innovative β-lactam/β-lactamase inhibitor compounds had proven to be an effective treatment option.

Antibiotic Susceptibility of Aerobic and Facultative Anaerobic Gram-Negative Rods in Hong Kong and Implications on Usefulness of Ceftazidime-Avibactam and Ceftolozane-Tazobactam.

Due to the increasing resistance of aerobic and facultative anaerobic Gram-negative rods, ceftazidime-avibactam and ceftolozane-tazobactam have been launched in the market in the last few years. In this study, we analyzed the susceptibility pattern of the major aerobic and facultative anaerobic Gram-negative rods in Hong Kong for ceftazidime-avibactam, ceftolozane-tazobactam, four other broad-spectrum antibiotics commonly used in Hong Kong and colistin. For 300 isolates collected from January to December 2021, non-ESBL-producing Enterobacterales, ESBL-producing Enterobacterales and Pseudomonas aeruginosa were highly susceptible to ceftazidime-avibactam (all 100%) and ceftolozane-tazobactam (98.7%, 99.7% and 94.3%). For 32 archived ESBL-producing Klebsiella pneumoniae isolates collected between January 2014 and March 2023, all were susceptible to ceftazidime-avibactam and ceftolozane-tazobactam. For 101 archived carbapenemase-producing Enterobacterales, their susceptibilities to ceftazidime-avibactam and ceftolozane-tazobactam varied depending on the type of carbapenemase produced. Both had high activities against OXA-producing strains (97.1% and 76.5%, respectively) but were 100% resistant for NDM-producing and NDM+OXA-producing strains. All KPC-producing strains were susceptible to ceftazidime-avibactam but resistant to ceftolozane-tazobactam. Ceftazidime-avibactam and ceftolozane-tazobactam are good alternatives for the management of infections caused by ESBL-producing Enterobacterales and selective strains of carbapenemase-producing Enterobacterales in Hong Kong.

Metallo-β-lactamases producing Pseudomonas aeruginosa and the molecular mechanism of drug resistance variants

BackgroundMetallo-β-lactamases (MBLs) type carbapenemases are produced by pathogenic Pseudomonas spp. and exhibit carbapenemase activity. The study will look into drug resistance and the molecular mechanisms of drug-resistant Pseudomonas aeruginosa variants. MethodsA total of 74 P. aeruginosa strains were isolated from urine, pus, sputum, blood, throat swab, Foley’s catheter, and nasal swab. The isolates were screened for antibiotic susceptibility and the identified MDR strains were further tested for β-lactamase production. Multiplex PCR was used to identify the presence of mcr-1 and blaNDM-1 genes in MDR organisms. The minimum inhibitory concentration for ceftazidime and colistin was also determined, in addition to the biofilm inhibitor activity. Confocal microscopy was used to determine the production of biofilms. ResultsThe isolated P. aeruginosa strains exhibited antibiotic resistance to aminoglycosides (amikacin and gentamicin). Fifty three percent of the isolated P. aeruginosa strains produced metallo-β-lactamase and the remaining isolates were non-metallo-β-lactamase type (46.8 %) (p < 0.0001). The MIC value of β-lactamase producers against colistin ranges from 0.5 µg/mL to 6 µg/mL. The MDR bacteria exhibited mcr-1 and blaNDM−1 genes. The MDR P. aeruginosa strain treated with colistin and ceftazidime inhibited initial biofilm formation. These combination of antibiotics effectively prevented initial biofilm development than individual antibiotics (p < 0.001). ConclusionsThe current analysis detected MDR among P. aeruginosa isolates that carried drug-resistant genes.

Modification of carbapenemase inhibition test and comparison of its performance with NG-Test CARBA 5 for detection of carbapenemase-producing enterobacterales.

Adequately and accurately identifying carbapenemase-producing Enterobacterales (CPE) is vital for selecting appropriate antimicrobial therapy and implementing effective infection control measures. This study aims to optimize the phenotypic detection method of carbapenemase for routine diagnostics in clinical microbiology laboratories. Carbapenemase genes in 2665 non-duplicate carbapenem-resistant Enterobacterales (CRE) clinical strains collected from various regions of China were confirmed through whole-genome sequencing (WGS). The carbapenemase inhibition test (CIT) was conducted and interpreted using different methods and breakpoints, then compared with the NG-Test CARBA 5 for carbapenemase detection. The diagnostic performance of the CIT method was optimal when the carbapenemase types were determined by comparing the inhibition zone diameters of the imipenem disc with 3-aminophenylboronic acid (APB) plus ethylenediaminetetraacetic acid (EDTA) to those of the imipenem disc with either APB or EDTA alone, with a breakpoint of 4mm. The overall sensitivities of the current CIT, the modified CIT and NG-Test CARBA 5 were 91.4%, 94.9% and 99.9%, respectively. For detecting isolates co-producing Klebsiella pneumoniae carbapenemase (KPC) and metallo-β-lactamases (MBLs), the modified CIT method had higher sensitivity than the current method (70.0%vs. 53.3%), though this difference was not statistically significant (p=0.063). The NG-Test CARBA 5 showed excellent performance for multi-carbapenemases diagnosis, with sensitivity and specificity of 97.1% and 100%, respectively. Optimizing and standardizing the CIT method for clinical use is necessary. It has certain advantages in diagnosing multi-carbapenemase and rare carbapenemase production. However, for identifying common carbapenemase types, the NG-Test CARBA 5 demonstrated superior performance.

Improved Blue Carba test and Carba NP test for detection and classification of major Class A and B carbapenemases, including dual producers, among Gram-negative bacilli.

Rapid and accurate identification of carbapenemase-producing organisms is of vital importance in guiding appropriate clinical antibiotic treatments and curbing their transmission. The emergence of negative bacilli carrying multiple carbapenemase combinations during and after the severe acute respiratory syndrome coronavirus 2 pandemic has posed a challenge to the conventional biochemical tests typically used to determine the specific carbapenemase type in the isolated strains. Several initiatives have aimed to enhance colorimetric methods, enabling them to independently identify the presence of Class A or Class B carbapenemases. Notably, no previous efforts have been made to distinguish both classes simultaneously. Additionally, these modifications have struggled to differentiate between carriers of multiple carbapenemases, a common occurrence in many Latin American countries. In this study, we introduced specific Class A and Class B carbapenemase inhibitors into the Blue Carba test (BCT) and Carba NP-direct (CNP) colorimetric assays to identify the type of carbapenemase, even in cases of multiple carbapenemase producers within these classes. These updated assays demonstrated exceptional sensitivity and specificity (≥ 90%) all within a rapid turnaround time of under 2 hours, typically completed in just 45 minutes. These in-house enhancements to the BCT and CNP assays present a rapid, straightforward, and cost-effective approach to determining the primary carbapenemase classes. They could serve as a viable alternative to molecular biology or immuno-chromatography techniques, acting as an initial diagnostic step in the process.

Minimum inhibitory concentrations of aztreonam-avibactam, ceftazidime-avibactam and meropenem in clinical isolates of Klebsiella pneumoniae harboring carbapenemase genes.

This study was aimed at understanding the distributions of the MICs (minimum inhibitory concentrations) of aztreonam-avibactam, ceftazidime-avibactam and meropenem with respect to Klebsiella pneumoniae isolates producing different types of carbapenemases and their combinations. K. pneumoniae isolates were collected between 2019 and 2022 from 37 hospitals. PCR was used to screen for blaKPC-, blaNDM- and blaOXA-48-like genes. MICs were determined by the broth microdilution method for meropenem, aztreonam-avibactam and ceftazidime-avibactam at a constant avibactam concentration of 4 mg l-1. MIC distributions were analyzed for groups of isolates based on the identified carbapenemases including their combinations. The AZT/AVI MIC50 and MIC90 for all NDM-positive isolates were 0.25 and 0.5, respectively, and for serine-carbapenemase-only producers, they were 0.25 and 1 mg l-1, respectively. The CZD/AVI MIC50 and MIC90 values for serine-carbapenemase-only producers were 1 and 4 mg l-1, respectively. The AZT/AVI MIC50 and MIC90 values for co-producers and single carbapenemase producers were the same (i.e., 0.25 and 1 mg l-1, respectively). The total proportion of meropenem-susceptible isolates (≤8 mg l-1) among all the carbapenemase producers was 25.1% (31.1% among single-carbapenemase producers and 9.2% among co-producers). The results support the use of aztreonam-avibactam for the empirical treatment of infections caused by any carbapenemase producers.

Epidemiological and clinical characteristics of patients with carbapenem-resistant Enterobacterales in a university hospital of Colombia: Enzyme coproductions in rise

The distribution of carbapenemases in Carbapenem-Resistant Enterobacterales (CRE) has recently undergone a change in our region. According to the Colombian National Institute of Health, there is an increasing prevalence of NDM and NDM-KPC co-producing strains. We carried-out an ambispective cohort study of adult inpatients from Hospital Universitario San Ignacio (2021–2023), infected or colonized with CRE, in which carbapenemases immunochromatographic assay was performed. Out of the 150 patients included in the study, 71.3 % presented with an infection, and carbapenemases were detected in 92.7 % of these cases. Among them, KPC predominated (54 %), while 16.7 % demonstrated enzyme coproductions, mainly KPC-NDM. CRE infected patients had an 18.7 % 30-days mortality, but we could not demonstrate an association between type of carbapenemase and mortality rate (p = 0.82). Logistic regression analysis suggested that ICU admission was independently correlated to fatality (OR 5.08; CI 1.68–16.01). NDM and KPC-NDM presence in CRE poses a public health threat and a therapeutic challenge, with unknown mortality differences according to the carbapenemases pattern. Nevertheless, there was not an association between enzyme type and mortality.

Detection of Carbapenemase Production in Klebsiella pneumoniae Isolates by Rapid Carbapenemase Detection Method in an OXA-48 Endemic Region

Background: Carbapenem resistance is one of the major global public health threats caused by the overuse or misuse of carbapenems, which are often the last resort in treating multidrug-resistant infections. Today, timely detection of carbapenemase-producing organisms is critical for infection control. Therefore, rapid and accurate detection of carbapenemases using a cost-effective method with high sensitivity and specificity is essential to guide appropriate treatment and prevent further spread. Objectives: This study aimed to evaluate the sensitivity of the newly developed rapid carbapenemase detection method (rCDM) with isolates whose resistance genes were determined by molecular methods and to compare it with the modified carbapenem inactivation method (mCIM), whose diagnostic performance characteristics are well-defined. Methods: A total of 130 Klebsiella pneumoniae isolates (65 carbapenem-susceptible, 65 carbapenem-resistant) from various clinical specimens were included in the study. Identification and antibiotic susceptibility testing of the isolates was performed using the BD Phoenix™ automated system. Phenotypic carbapenemase detection methods, mCIM and rCDM, were studied for all carbapenem-sensitive and carbapenem-resistant isolates with known resistance genes. Results: Among carbapenem-resistant K. pneumoniae isolates, blaOXA-48 was the most frequently detected resistance gene (78%). Using PCR results as the reference method, the sensitivity of rCDM was 100%. When rCDM and mCIM results were compared, both phenotypic methods were 100% compatible, with no statistically significant difference observed. Conclusions: Rapid carbapenemase detection method is a rapid and reliable phenotypic carbapenemase detection method with high sensitivity. Its ease of use and interpretation make it suitable for use in laboratories with limited resources. Additionally, its excellent performance in detecting common carbapenemase types and high consistency with mCIM support its utility in various laboratory settings without requiring additional materials or technical infrastructure.

Urinary Tract Infections with Carbapenem-Resistant Klebsiella pneumoniae in a Urology Clinic-A Case-Control Study.

The aim of our study was to analyze the factors associated with the increased risk of urinary tract infection (UTI) with carbapenem-resistant (CR) Klebsiella pneumoniae (Kpn) and the antibiotic resistance spectrum of the strains in patients. As secondary objectives, we elaborated the profile of these patients and the incidence of different types of carbapenemases. We conducted a retrospective case-control study in which we compared a group of 62 patients with urinary tract infections with CR Kpn with a control group consisting of 136 patients with urinary tract infections with multidrug-resistant (MDR), but carbapenem-sensitive (CS), Kpn, who were hospitalized between 1 January 2022 and 31 March 2024. Compared to patients with urinary tract infections with CS Kpn, patients with urinary tract infections with CR Kpn were preponderant in rural areas (62.9% vs. 47.1%, p = 0.038) and more frequently had an upper urinary tract infection (69.4% vs. 36.8%, p < 0.01). Among the risk factors examined, patients in the study group had a higher presence of urinary catheters inserted for up to one month (50% vs. 34.6%, p = 0.03), rate of hospitalization in the last 180 days (96.8% vs. 69.9%, p < 0.01) and incidence of antibiotic therapy in the last 180 days (100% vs. 64.7%, p < 0.01). They also had a higher rate of carbapenem treatment in the last 180 days (8.1% vs. 0%, p < 0.01). Patients in the study group had a broader spectrum of resistance to all antibiotics tested (p < 0.01), with the exception of sulfamethoxazole-trimethoprim, where the resistance rate was similar in both groups (80.6% vs. 67.6%, p = 0.059). In the multivariate analysis, transfer from other hospitals (OR = 3.51, 95% and CI: 1.430-8.629) and treatment with carbapenems in the last 180 days (OR = 11.779 and 95% CI: 1.274-108.952) were factors associated with an increased risk of disease compared to the control group. The presence of carbapenemases was observed in all patients with CR Kpn, in the order of frequency New Delhi metallo-ß-lactamase (NDM) (52.2%), Klebsiella pneumoniae carbapenemase (KPC) (32.6%), and carbapenem-hydrolyzing oxacillinase (Oxa-48) (15.2%). The environment of origin and previous treatment with carbapenems appear to be the factors associated with an increased risk of urinary tract infection with CR Kpn compared to patients with urinary tract infections with CS Kpn. CR Kpn exhibits a broad spectrum of antibiotic resistance, among which is resistance to carbapenem antibiotics.

Plasmid genomic epidemiology of carbapenem-hydrolysing class D β-lactamase (CDHL)-producing Enterobacterales in Canada, 2010-2021.

Carbapenems are last-resort antibiotics for treatment of infections caused by multidrug-resistant Enterobacterales, but carbapenem resistance is a rising global threat due to the acquisition of carbapenemase genes. Oxacillinase-48 (bla OXA-48)-type carbapenemases are increasing in abundance in Canada and elsewhere; these genes are frequently found on mobile genetic elements and are associated with specific transposons. This means that alongside clonal dissemination, bla OXA-48-type genes can spread through plasmid-mediated horizontal gene transfer. We applied whole genome sequencing to characterize 249 bla OXA-48-type-producing Enterobacterales isolates collected by the Canadian Nosocomial Infection Surveillance Program from 2010 to 2021. Using a combination of short- and long-read sequencing, we obtained 70 complete and circular bla OXA-48-type-encoding plasmids. Using MOB-suite, four major plasmids clustered were identified, and we further estimated a plasmid cluster for 91.9 % (147/160) of incomplete bla OXA-48-type-encoding contigs. We identified different patterns of carbapenemase mobilization across Canada, including horizontal transmission of bla OXA-181/IncX3 plasmids (75/249, 30.1 %) and bla OXA-48/IncL/M plasmids (47/249, 18.9 %), and both horizontal transmission and clonal transmission of bla OXA-232 for Klebsiella pneumoniae ST231 on ColE2-type/ColKP3 plasmids (25/249, 10.0 %). Our findings highlight the diversity of OXA-48-type plasmids and indicate that multiple plasmid clusters and clonal transmission have contributed to bla OXA-48-type spread and persistence in Canada.

Prevalence and antimicrobial mechanism of carbapenem-resistant Klebsiella pneumoniae and its molecular properties

BackgroundThe carbapenem-resistant Klebsiella pneumoniae poses a serious threat to public health because carbapenems are used as a final resort to treat K. pneumoniae-mediated infections in humans. MethodsSamples were collected from various clinical specimens including skin swabs, anal swabs, wound swabs, oral swabs, and sputum by the standard method and K. pneumoniae strains were isolated. Biofilm-forming characters were determined. The antibiotic resistance pattern was analyzed by the Kirby–Bauer disk diffusion method. Carbapenem resistance properties were tested using the imipenem and meropenem antibiotics. The carbapenemase genes (blaKPC and blaNDM) were determined. ResultsA total of 11 cephalosporin-resistant K. pneumoniae (CRKP) strains were isolated from the samples. The screened CRKP strains exhibited multi-drug resistance and non-susceptible to imipenem, ceftazidime, piperacillin, ceftriaxone, cefazolin, ampicillin, aztreonam, and cefotetan antibiotics. A total of 79.4 % K. pneumoniae isolates showed positive results on String-forming test. About 82.1 % of isolates showed mucoid colonies and 59 % of K. pneumoniae strains formed biofilm (p < 0.01). Out of 11 isolates, four strains exhibited Klebsiella pneumoniae carbapenemase type, three strains produced metallo-β-lactamases (MBL), and four strains exhibited as carbapenemase types. A total of 63.5 % Klebsiella pneumoniae carbapenemase-producing strains showed very low MIC value (<0.05 mg/L). ConclusionsDrug-resistance K. pneumoniae was isolated from clinical specimens that were screened. The continuous monitoring of drug-resistance genes are required to make policy decisions.

Comparison of ERIC carbapenem-resistant Enterobacteriaceae test, BD Phoenix CPO detect panel, and NG-test CARBA 5 for the detection of main carbapenemase types of carbapenem-resistant Enterobacterales

BackgroundThis study aimed to assess the performance of three commercial panels, the ERIC Carbapenem-Resistant Enterobacteriaceae Test (ERIC CRE test), the NG-Test CARBA 5 (NG CARBA 5), and the BD Phoenix CPO Detect Panel (CPO panel), for the detection of main types of carbapenemases among carbapenem-resistant Enterobacterales (CRE). MethodsWe collected 502 isolates of carbapenem-resistant Enterobacterales (CRE) demonstrating intermediate or resistant profiles to at least one carbapenem antibiotic (ertapenem, imipenem, meropenem, or doripenem). Carbapenemase genes and their specific types were identified through multiplex PCR and sequencing methods. Subsequently, the ERIC CRE test, CPO panel, and NG CARBA 5 assay were conducted on these isolates, and the results were compared with those obtained from multiplex PCR. ResultsThe results indicated that the ERIC CRE test exhibited an overall sensitivity and specificity of 98.1% and 93.6%, respectively, which were comparable to 99.1% and 90.6% for the NG CARBA 5. However, the CPO panel demonstrated a sensitivity of only 56.2% in identifying Ambler classes, exhibiting the poorest sensitivity for class A. Moreover, while the ERIC CRE test outperformed the NG CARBA 5 in identifying multi-gene isolates with multiple carbapenemase-encoding genes, the CPO panel failed to accurately classify these isolates. ConclusionsOur findings support the utilization of the ERIC CRE test as one of the methods for detecting carbapenemases in clinical laboratories. Nonetheless, further optimization is imperative for the CPO panel to enhance its accuracy in determining carbapenemase classification and address limitations in detecting multi-gene isolates.

Carbapenem-Resistant NDM and OXA-48-like Producing K. pneumoniae: From Menacing Superbug to a Mundane Bacteria; A Retrospective Study in a Romanian Tertiary Hospital.

Carbapenem-resistant Klebsiella pneumoniae (Cr-Kpn) is becoming a growing public health problem through the failure of adequate treatment. This study's objectives are to describe the sources of Cr-Kpn in our hospital over 22 months, associating factors with the outcome of Cr-Kpn-positive patients, especially those with NDM+OXA-48-like (New Delhi Metallo-β-Lactamase and oxacillinase-48), and the effectiveness of the treatments used. A retrospective observational cohort study including all hospitalized patients with Cr-Kpn isolates. We reported data as percentages and identified independent predictors for mortality over hospital time through multivariate analysis. The main type of carbapenemases identified were NDM+OXA-48-like (49.4%). The statistical analysis identified that diabetes and co-infections with the Gram-negative, non-urinary sites of infection were factors of unfavorable evolution. The Cox regression model identified factors associated with a poor outcome: ICU admission (HR of 2.38), previous medical wards transition (HR of 4.69), and carbapenemase type NDM (HR of 5.98). We did not find the superiority of an antibiotic regimen, especially in the case of NDM+OXA-48-like. The increase in the incidence of Cr-Kpn infections, especially with NDM+OXA-48-like pathogens, requires a paradigm shift in both the treatment of infected patients and the control of the spread of these pathogens, which calls for a change in public health policy regarding the use of antibiotics and the pursuit of a One Health approach.

Detection of Carbapenem Resistance Using the Genotypic and Phenotypic Methods in Klebsiella pneumoniae

Aim: This study aimed to detect the carbapenem resistance of the Klebsiella pneumoniae strains, isolated from clinical specimens with genotypic and phenotypic methods. Material and Methods: A total of 87 Klebsiella pneumoniae strains whose carbapenem resistance was determined by disc diffusion method were included in the study. Carbapenemase was investigated using the combined disk method and polymerase chain reaction (PCR). Results: The evaluation of the PCR results demonstrated that OXA was detected in 60 (68.9%) samples, NDM was detected in 20 (22.9%), OXA + NDM in 5 (5.7%), and KPC was detected in 1 (1.1%) out of 87 clinical samples. Carbapenemase was not detected in one specimen with the PCR method. The results were found compatible with the combined disc test results for all isolates which were detected as only OXA, NDM, and KPC type carbapenemase positive. In 5 (5.7%) strains in which the co-existence of NDM and OXA type carbapenemases was detected by PCR, the combined disc method detected only OXA type carbapenemase. Conclusion: The combined disk method is inadequate in the presence of strains that have multiple carbapenemases, and also have OXA which is the most frequently detected carbapenemase in our hospital. EUCAST recommends verification by other methods in the presence of OXA-48. Genotypic methods can be used for confirmation testing. The detections of strains with NDM, multiple carbapenemases, and the first detection of KPC were striking in the study. Monitoring the spread of these strains in the hospital will be necessary for infection control.

Enterobacteriaceae (Order Enterobacterales), &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt; and &lt;i&gt;Acinetobacter spp&lt;/i&gt; Carbapenemase producers isolated from patients hospitalized in ICU during the COVID-19 pandemic, 2020.

The present study aimed to determine the type of carbapenemase present in Enterobactales, Pseudomonas aeruginosa and Acinetobacter spp. isolated from patients hospitalized in the ICU of Hospital IV "Víctor Lazarte Echegaray", during the COVID-19 pan-demic. In this descriptive, observational and cross-sectional study, biological samples were collected from patients hospitalized in the ICU, processed in the Microbiology Diagnostic Laboratory of Hospital IV "Víctor Lazarte Echegaray". Antimicrobial identification and sensitivity were performed with the AutoScan-4 automated system, and the RESISIT-3 O.K.N K-SET cassettes were used to identify the type of carbapenemase. Of a total of 129 cultures, 58,9% were found to be resistant to at least one carbapenem (Meropenem or Imipenem), of which 59,2% were Enterobacterales, 26,3% were Pseudomonas aeruginosa and 14.5% were% Acinetobacter spp; when evaluating the presence of carbapenemases, it was found that 66 cultures (51,2%) presented carbapenemases; of which 28 are positive to KPC (42,4%), 25 to NDM (37,9%) and 13 to OXA-48 (19,7%). It is concluded that the presence of carbapenemasas in microbial cultures isolated from patient hospitalized I the intensive care unit exceeds the 30% reported in 2019 in South America.

Molecular characteristics of carbapenem-resistant Enterobacteriaceae isolated from two tertiary hospitals in the Philippines

Species belonging to the family Enterobacteriaceae cause infections that result in clinically significant morbidity and mortality. These organisms have been shown to harbor multiple mechanisms of resistance to antibiotics, including carbapenems. Molecular characterization of resistance to carbapenems is critical since few effective therapeutic options exist, most of which are difficult to access in resource-limited settings. One hundred twenty-six Enterobacteriaceae isolates with phenotypic resistance to at least one carbapenem were collected from two tertiary hospitals in the Philippines from 2014 to 2016. The organisms were screened for the presence of genes coding for carbapenemases and extended spectrum β-lactamases (ESBLs) using multiplex PCR. Multilocus sequence typing (MLST) was performed for Klebsiella pneumoniae, Escherichia coli and Enterobacter cloacae complex. The majority of the carbapenem-resistant isolates were K. pneumoniae (70/126, 55.5%) followed by K. ozaenae (19/126, 15%). Amplification of genes conferring carbapenem resistance showed blaNDM in 89 isolates (70%), followed by blaKPC in 21 isolates (16%). MLST revealed the presence of different clones including ST16 (5/70), ST17 (3/70) and ST147 (9/70) in K. pneumoniae, and ST10 (3/13) in E. coli. This study shows that blaNDM is the most prevalent carbapenemase type among carbapenem-resistant Enterobacteriaceae (CRE). Presence of carbapenemase gene and multiple ESBLs in the same organism pose a risk in the clinical setting since it could confer higher antibiotic resistance. This high rate of resistance poses the risk of spread since it can be transferred by mobile genetic elements. This study illustrates the acute need for effective therapeutic and hospital infection controls against CREs.

Pseudomonas aeruginosa with double carbapenemase type IMP and KPC in a pediatric hospital in Lima, Peru

Sr. Editor, Pseudomonas aeruginosa es un patógeno oportunista, causante de infecciones asociadas a la atención en salud (IAAS). El tratamiento es limitado debido a la resistencia intrínseca de la especie y a la resistencia adquirida, dejando a los carbapenémicos como la única alternativa terapéutica para aislamientos multirresistentes (1).