Ceftaroline Susceptibility Research Articles

In Vitro Activity of Ceftaroline against Staphylococcus aureus Isolated in 2012 from Asia-Pacific Countries as Part of the AWARE Surveillance Program.

Ceftaroline, the active metabolite of the prodrug ceftaroline-fosamil, is an advanced-generation cephalosporin with activity against methicillin-resistant Staphylococcus aureus (MRSA). This investigation provides in vitro susceptibility data for ceftaroline against 1,971 S. aureus isolates collected in 2012 from seven countries (26 centers) in the Asia-Pacific region as part of the Assessing Worldwide Antimicrobial Resistance and Evaluation (AWARE) program. Broth microdilution as recommended by the CLSI was used to determine susceptibility. In all, 62% of the isolates studied were MRSA, and the ceftaroline MIC90 for all S. aureus isolates was 2 μg/ml (interpretive criteria: susceptible, ≤1 μg/ml). The overall ceftaroline susceptibility rate for S. aureus was 86.9%, with 100% of methicillin-sensitive S. aureus isolates and 78.8% of MRSA isolates susceptible to this agent. The highest percentages of ceftaroline-nonsusceptible MRSA isolates came from China (47.6%), all of which showed intermediate susceptibility, and Thailand (37.1%), where over half (52.8%) of isolates were resistant to ceftaroline (MIC, 4 μg/ml). Thirty-eight ceftaroline-nonsusceptible isolates (MIC values of 2 to 4 μg/ml) were selected for molecular characterization. Among the isolates analyzed, sequence type 5 (ST-5) was the most common sequence type encountered; however, all isolates analyzed from Thailand were ST-228. Penicillin-binding protein 2a (PBP2a) substitution patterns varied by country, but all isolates from Thailand had the Glu239Lys substitution, and 12 of these also carried an additional Glu447Lys substitution. Ceftaroline-fosamil is a useful addition to the antimicrobial agents that can be used to treat S. aureus infections. However, with the capability of this species to develop resistance to new agents, it is important to recognize and monitor regional differences in trends as they emerge.

Potential of Staphylococcus aureus isolates carrying different PBP2a alleles to develop resistance to ceftaroline.

Infections caused by MRSA continue to cause significant morbidity worldwide. Ceftaroline (the active metabolite of the prodrug ceftaroline fosamil) is a cephalosporin that possesses activity against MRSA due to its having high affinity for PBP2a while maintaining activity against the other essential PBPs. PBP2a sequence variations, including some outside of the transpeptidase binding pocket, impact ceftaroline susceptibility. This study evaluated the potential of ceftaroline to select for resistant Staphylococcus aureus clones in isolates containing a variety of PBP2a alleles and with a range of ceftaroline MIC values from different MLST lineages. Direct resistance selection experiments were performed by plating 20 S. aureus isolates (18 MRSA and 2 MSSA) on agar plates containing increasing concentrations of ceftaroline. Colonies that emerged were tested by standard broth microdilution for changes in ceftaroline susceptibility and genetically characterized. The frequency of spontaneous resistance to ceftaroline was low for all isolates and, although resistant variants were not obtained on plates containing ≥4-fold the MIC of ceftaroline, six MRSA isolates had a small number of colonies emerge on plates containing 2-fold the MIC of ceftaroline and had a 2- to 8-fold elevation of the ceftaroline MIC, while also impacting the MIC of methicillin compared with the parental isolate. Additional PBP2a mutations located in the ceftaroline-binding pocket, Y446N or A601S, were observed in several of the resistant isolates. These studies demonstrate that there is a low risk of generating ceftaroline-resistant MRSA isolates, which appears independent of any pre-existing variation in the PBP2a protein sequence or initial ceftaroline MIC.

Reduced In Vitro Activity of Ceftaroline by Etest among Clonal Complex 239 Methicillin-Resistant Staphylococcus aureus Clinical Strains from Australia.

A total of 421 methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates were tested for ceftaroline susceptibility by Etest (bioMérieux). A multidrug resistant phenotype was found in 40.9%, and clonal complex 239 (CC239) was found in 33.5%. Ceftaroline nonsusceptibility (MIC, >1.0 μg/ml) was 16.9% overall. Nonsusceptibility was significantly higher in CC239 (41.1%, 58/141) and in isolates with a multidrug resistant phenotype (35.5%, 61/172) compared with comparators (P < 0.0001). Nonsusceptibility of common multidrug resistant MRSA clones limits the empirical use of ceftaroline for these infections.

Molecular characterization of MRSA isolates bracketing the current EUCAST ceftaroline-susceptible breakpoint for Staphylococcus aureus: the role of PBP2a in the activity of ceftaroline.

The objectives of this study were to characterize contemporary MRSA isolates and understand the prevalence and impact of sequence variability in PBP2a on ceftaroline susceptibility. A total of 184 MRSA isolates collected from 28 countries were collected and characterized. WT PBP2a proteins were found in MRSA distributed evenly over the ceftaroline MIC range of 0.5-2 mg/L (n=56). PBP2a variations found in 124 isolates fell into two categories: (i) 12 isolates contained a substitution in the transpeptidase pocket located in the penicillin-binding domain and exhibited significantly decreased ceftaroline susceptibility (typically 8 mg/L); and (ii) isolates with substitutions in the non-penicillin-binding domain (nPBD) in a region proposed to be functionally important for cell wall biogenesis. The majority (71%) of isolates containing only nPBD variations were inhibited by 2 mg/L ceftaroline, 23% by ≤1 mg/L and 6% by 4 mg/L. These data suggest that the WT MRSA distribution extends beyond the current EUCAST and CLSI susceptible breakpoints and includes isolates inhibited by 2 mg/L ceftaroline. SCCmec type IV was the predominant type in the ceftaroline-susceptible population (68%), whereas it only represented 6% of the non-susceptible population. The variations of MLST lineages were fewer among the non-susceptible group. This study suggests that MRSA populations with a WT PBP2a and those with nPBD variations overlap significantly and that PBP2a sequence-independent factors contribute to ceftaroline susceptibility. Whereas characterization of isolates with a ceftaroline MIC of 2 mg/L enriched for isolates with nPBD variations, it was not a discrete population. In contrast, the rare isolates containing a substitution in the transpeptidase-binding pocket were readily differentiated.

Missense mutations in PBP2A Affecting ceftaroline susceptibility detected in epidemic hospital-acquired methicillin-resistant Staphylococcus aureus clonotypes ST228 and ST247 in Western Switzerland archived since 1998.

The development and maintenance of an arsenal of antibiotics is a major health care challenge. Ceftaroline is a new cephalosporin with activity against methicillin-resistant Staphylococcus aureus (MRSA); however, no reports concerning MRSA ceftaroline susceptibility have been reported in Switzerland. We tested the in vitro activity of ceftaroline against an archived set of 60 MRSA strains from the University Hospital of Geneva collected from 1994 to 2003. Our results surprisingly revealed ceftaroline-resistant strains (MIC, >1 μg/ml in 40/60 strains; EUCAST breakpoints, susceptible [S], ≤1 μg/ml; resistant [R], >1 μg/ml) were present from 1998 to 2003. The detected resistant strains predominantly belonged to sequence type 228 (ST228) (South German clonotype) but also to ST247 (Iberian clonotype). A sequence analysis of these strains revealed missense mutations in the penicillin-binding protein 2A (PBP2A) allosteric domain (N146K or E239K and N146K-E150K-G246E). The majority of our ST228 PBP2A mutations (N146K or E150K) were distinct from ST228 PBP2A allosteric domain mutations (primarily E239K) recently described for MRSA strains collected in Thailand and Spain during the 2010 Assessing Worldwide Antimicrobial Resistance Evaluation (AWARE) global surveillance program. We also found that similar allosteric domain PBP2A mutations (N146K) correlated with ceftaroline resistance in an independent external ST228 MRSA set obtained from the nearby University Hospital of Lausanne, Lausanne, Switzerland, collected from 2003 to 2008. Thus, ceftaroline resistance was observed in our archived strains (including two examples of an MIC of 4 µg/ml for the Iberian ST247 clonotype with the triple mutation N146K/E150K/G246E), at least as far back as 1998, considerably predating the commercial introduction of ceftaroline. Our results reinforce the notion that unknown parameters can potentially exert selective pressure on PBP2A that can subsequently modulate ceftaroline resistance.

Development of EUCAST zone diameter breakpoints and quality control range for Staphylococcus aureus with ceftaroline 5-μg disk.

This ceftaroline MIC/disk comparison study for Staphylococcus aureus was performed for the purpose of establishing EUCAST zone diameter breakpoints. Ceftaroline susceptibility for a challenge set of 70 methicillin resistant- and 30 methicillin susceptible-S. aureus was determined by 5-μg disk diffusion and broth microdilution methods. Seventeen isolates were retested by disk and MIC, and the remaining 83 isolates were retested by MIC. Molecular testing was performed on 19 isolates with borderline susceptible ceftaroline MIC results to assess any differences in mecA and epidemiological correlation. An additional set of 101 consecutive clinical S. aureus isolates were tested using the 5-μg disk. S. aureus ATCC 29213 was tested by multiple sites and media for QC range determination. Replicate MIC results were within ±1 doubling dilution, with tendency for slightly lower repeat MICs, and there was minimal variation in replicate zone results. Based on susceptible breakpoints for MIC of ≤1 mcg/mL and for disk of >20 mm, there was 100 % categorical agreement for 30 MSSA and 92 % categorical agreement for 70 MRSA. There were no common MLST or PBP changes for strains with MICs of 1 and 2 mcg/mL. All ceftaroline disk results for the consecutively collected isolates were >20 mm. EUCAST selected the ceftaroline 5-μg disk breakpoint of Susceptible ≥20, Resistant <20 mm because it correlated best with the MIC breakpoint of Susceptible ≤1, Resistant >1 mg/L. A ceftaroline 5-μg disk QC range for S. aureus ATCC 29213 of 24-30 mm was also established by EUCAST.

Observation of "seesaw effect" with vancomycin, teicoplanin, daptomycin and ceftaroline in 150 unique MRSA strains.

IntroductionVancomycin (VAN) failures associated with the treatment of complicated methicillin-resistant Staphylococcus aureus (MRSA) infections have been well described. The reported “seesaw effect” demonstrates improved β-lactam activity when VAN and/or daptomycin (DAP) susceptibility decreases. However, there are minimal data comparing ceftaroline (CPT) susceptibility with these agents or teicoplanin (TEI). Therefore, to further explore the seesaw effect, we evaluated the relationship between CPT and VAN, TEI, and DAP susceptibilities.MethodsOne hundred and fifty clinical MRSA isolates from the Anti-Infective Research Laboratory (Detroit, MI, USA) from 2008 to 2012 were analyzed. VAN, TEI, DAP and CPT minimum inhibitory concentrations (MIC) were determined via Etest methodology. MIC50 and MIC90 were calculated for each antibiotic. Additionally, four isogenic strain pairs were randomly selected for evaluation by time–kill methodology for the potential of enhanced killing by CPT as MICs increased to VAN, TEI, and DAP.ResultsCPT MICs were inversely correlated with VAN, DAP, and TEI MICs with correlation coefficients of −0.535, −0.483, and −0.386, respectively (P ≤ 0.05). Comparison of the MIC relationship for glycopeptides and lipopeptides resulted in a positive correlation for all agent combinations. In time–kill evaluations, CPT demonstrated greater reductions in log10 colony-forming unit (CFU)/mL against mutant strains (3.73 ± 0.67) versus parents (2.79 ± 0.75) despite no change in CPT MIC (P = 0.112).ConclusionThis study demonstrated a marked “seesaw effect” whereby CPT displayed increased susceptibility as the VAN, DAP, and TEI MICs increased. Additionally, we observed a positive linear correlation between VAN, DAP, and TEI MICs for all agent combinations. Enhanced activity was noted with CPT in mutant strains versus the parent strains despite no change in MIC. Based upon the enhanced CPT activity observed against strains with decreased susceptibility to VAN, DAP and TEI, CPT may provide an option for infections with reduced susceptibility to glycopeptides or lipopeptides. Further evaluation is warranted to investigate the clinical implications of the seesaw effect.Electronic supplementary materialThe online version of this article (doi:10.1007/s40121-014-0023-0) contains supplementary material, which is available to authorized users.

Activities of Vancomycin, Ceftaroline, and Mupirocin against Staphylococcus aureus Isolates Collected in a 2011 National Surveillance Study in the United States

Forty-two medical centers from throughout the United States participating in a longitudinal surveillance program were asked to submit 100 consecutive Staphylococcus aureus isolates during July to December 2011. Susceptibility testing using CLSI broth microdilution and mecA detection by PCR analysis was performed on the 4,131 isolates collected. Methods employing Etest glycopeptide resistance detection (GRD; bioMérieux) and brain heart infusion agar containing 4 μg/ml vancomycin (BHIV) were used to screen methicillin-resistant S. aureus (MRSA) isolates for heterogeneous intermediate-level resistance to vancomycin (hVISA). Isolates with positive hVISA screen results were confirmed by population analysis profiling-area under the curve (PAP-AUC) determinations. The genetic relatedness of hVISA, ceftaroline-nonsusceptible, or high-level (HL) mupirocin resistance MRSA isolates was assessed by pulsed-field gel electrophoresis (PFGE). Among 2,093 MRSA isolates, the hVISA screen results were positive with 47 isolates by Etest GRD and 30 isolates by BHIV agar screen. Twenty-five of the GRD- or BHIV screen-positive isolates were confirmed as hVISA by PAP-AUC testing. Results of the current study were compared to results obtained from prior surveillance performed in 2009. The prevalence of hVISA among MRSA isolates was higher in 2011 than in 2009 (1.2% versus 0.4%, P = 0.003), especially for isolates with a vancomycin MIC of 2 (45.4% versus 14.3%, P = 0.01). The overall rate of ceftaroline susceptibility in the current study was 99.4% (one hVISA isolate had an intermediate ceftaroline MIC). HL mupirocin resistance increased from 2.2% in 2009 to 3.2% in 2011 (P = 0.006). Although overall rates of hVISA and HL mupirocin resistance are low, they have increased since 2009.

Interim susceptibility testing for ceftaroline, a new MRSA-active cephalosporin: selecting potent surrogate β-lactam markers to predict ceftaroline activity against clinically indicated species

Ceftaroline, the bio-active form of parenterally administered ceftaroline fosamil, is a unique broad-spectrum cephalosporin with in vitro and in vivo activity against methicillin-resistant Staphylococcus aureus and was approved for clinical use by the United States Food and Drug Administration in October 2010. In over a year since ceftaroline fosamil approval, no widely used commercial susceptibility test system has added this new compound to its product, therefore requiring use of alternative agar diffusion methods for clinical microbiology laboratories that want to test clinical isolates for ceftaroline susceptibility. An alternative strategy of applying a surrogate β-lactam class marker agent was assessed here, using results from 14,902 organisms (2008–2010) sampled in the USA. Very high and acceptable accuracy (≥99.75%) was observed for predicting ceftaroline susceptibility as follows: 1) use of imipenem or meropenem minimum inhibitory concentrations (MICs) at ≤8 μg/mL (susceptible and intermediate categories) when testing S. aureus; 2) use of ceftriaxone MIC at ≤2 μg/mL (susceptible and intermediate categories) when testing Streptococcus pneumoniae as well as other streptococci (S. pyogenes and S. agalactiae); and 3) use of ceftriaxone, or cefepime, or ceftazidime at ≤2 μg/mL (susceptible category) when testing Haemophilus influenzae. Only when testing indicated Enterobacteriaceae species using ceftriaxone susceptibility results did the ceftaroline-nonsusceptible errors increase (4.11%). These presented analyses offer a validated surrogate marker strategy for ceftaroline susceptibility testing, pending development and validation by the commonly used automated systems and agar diffusion commercial methods.

Ceftaroline: a Cephalosporin with Anti-MRSA Activity

Ceftaroline is a cephalosporin antibiotic with broad-spectrum activity against gram-positive and gram-negative bacteria, including contemporary resistant gram-positive phenotypes, such as methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant Streptococcus pneumoniae, and penicillin-resistant S. pneumoniae. Ceftaroline is administered as the prodrug ceftaroline fosamil, which is rapidly converted via plasma phosphatases to its bioactive metabolite, ceftaroline. Ceftaroline fosamil was approved in October 2010 by the U.S. Food and Drug Administration (FDA) and launched in March 2011 for the treatment of acute bacterial skin and skin structure infections and for the treatment of community-acquired bacterial pneumonia caused by designated susceptible isolates. The Clinical and Laboratory Standards Institute (CLSI) designated ceftaroline a member of a new subclass of β-lactam antimicrobials, cephalosporins with anti-MRSA activity, based on its unique spectrum of activity. At this time, no ceftaroline surrogate is recommended for in vitro susceptibility testing because no other FDA-approved β-lactam has a similar spectrum of activity, although studies to identify potential surrogates are in progress. Ceftaroline susceptibility testing can be performed using CLSI standardized dilution or diffusion techniques, and FDA susceptibility interpretive criteria for broth dilution MIC values and disk diffusion zone diameters are available. Commercial broth-based antimicrobial susceptibility tests for ceftaroline are in development. A ceftaroline Etest for MIC testing is also in development. Ceftaroline Kirby-Bauer disks for diffusion were recently approved by the FDA and have been validated against the reference broth microdilution method.

Comparative ceftaroline activity tested against pathogens associated with community-acquired pneumonia: results from an international surveillance study

To document the spectrum of activity of ceftaroline, the active form of the prodrug, ceftaroline fosamil, a new cephalosporin with anti-methicillin-resistant staphylococcal activity, against a surveillance collection of clinical isolates obtained from the USA and Europe during 2008-09. A selected group of species associated with community-acquired pneumonia (CAP; 6496 of 17 326 monitored strains) were tested for susceptibility in a central laboratory using CLSI broth microdilution methods. Organisms were sampled from 55 medical centres, 27 in the USA and 28 (12 countries) in Europe. Ceftaroline and comparator agents were tested and interpretations of MIC endpoints made by applying current CLSI (2010) and EUCAST (2010) breakpoint criteria. Against 1340 Streptococcus pneumoniae, ceftaroline inhibited all isolates at ≤0.5 mg/L (MIC(50/90), ≤0.008/0.12 mg/L) and was 8-fold more active than ceftriaxone (MIC(90), 1 mg/L; only 79.2% coverage at EUCAST breakpoint). Haemophilus influenzae (n = 584; MIC(50/90), ≤0.008/0.015 mg/L), Moraxella catarrhalis (n = 377; MIC(50/90), 0.03-0.06/0.12 mg/L) and Staphylococcus aureus (n = 590; MIC(50/90), 0.5/1 mg/L) were very susceptible to ceftaroline, regardless of β-lactamase production or multidrug resistance (MDR) patterns. The potency of ceftaroline against three species of Enterobacteriaceae (Escherichia coli, Klebsiella pneumoniae and Enterobacter cloacae) was similar to that of ceftriaxone, ceftazidime and piperacillin/tazobactam. Only modest differences in rates of ceftaroline susceptibility (breakpoint ≤2 mg/L) were noted with extended-spectrum β-lactamase-negative Enterobacteriaceae strains between the USA and Europe (97.9% versus 97.0% for E. coli). Ceftaroline, like ceftriaxone, was not active against ceftazidime-resistant E. coli (10.2%-26.2% susceptible at ≤2 mg/L) or K. pneumoniae (5.3%-11.2%). The ceftaroline surveillance for 2008-09 (USA and Europe) documented low MIC(50/90) values for S. aureus isolates at 0.5/1 and 0.25/1 mg/L, respectively. More importantly, ceftaroline MIC(90) results for S. pneumoniae (0.12 mg/L), H. influenzae (0.015 mg/L) and M. catarrhalis (0.12 mg/L) were very low, all MICs being ≤0.5 mg/L. Ceftaroline exhibited promising high potency and wide coverage against Gram-positive and -negative pathogens known to cause CAP, especially isolates of MDR pneumococci and methicillin-resistant S. aureus.