In vitro activity of fosfomycin alone and in combination against Staphylococcus aureus with reduced susceptibility or resistance to methicillin, vancomycin, daptomycin or linezolid.

OBJECTIVES: To evaluate the activity of fosfomycin against a group of MRSA strains, including isolates with reduced susceptibility or resistance to vancomycin, daptomycin, linezolid and ceftaroline and to determine the effect of combining various combinations of antimicrobial agents used in the therapy of serious Gram-positive infections. METHODS: Broth microdilution testing was used to determine the MICs of fosfomycin, vancomycin, daptomycin, linezolid, ceftaroline and cefazolin. Isolates were selected for further evaluations to determine in vitro synergy between fosfomycin and select antimicrobial agents using chequerboard broth microdilution testing. Fosfomycin was tested in combination with vancomycin, linezolid, daptomycin, ceftaroline and cefazolin. RESULTS: Fosfomycin maintained activity against 100% of strains of vancomycin-resistant Staphylococcus aureus (VRSA) and linezolid-resistant S. aureus (LRSA), 86% of VISA and 95% of daptomycin-resistant S. aureus (DRSA) strains. The combination of fosfomycin with ceftaroline consistently demonstrated synergy among all 18 isolates against the strains tested. The next most potent combination regimen was linezolid with fosfomycin, which demonstrated synergy in 16 of the 18 strains. Daptomycin demonstrated synergy in only 7 of the 18 strains tested when combined with fosfomycin. Cefazolin demonstrated synergy in 6 of 6 strains and vancomycin demonstrated no interaction in 6 of 6 strains tested. CONCLUSIONS: Fosfomycin demonstrated excellent activity against MRSA as well as isolates with resistance or reduced activity to other anti-MRSA drugs including vancomycin, daptomycin and linezolid. When combined with linezolid or daptomycin, fosfomycin demonstrated synergy for all or most strains tested. Thus, these combinations may have potential clinical utility when treating patients with serious infections caused by MRSA.

Delafloxacin activity against Staphylococcus aureus with reduced susceptibility or resistance to methicillin, vancomycin, daptomycin or linezolid.

BACKGROUND: Delafloxacin is a recently approved anionic fluoroquinolone antibiotic with broad-spectrum activity against Gram-positive and Gram-negative organisms. The drug has been approved for patients with acute bacterial skin and skin structure infections including those caused by MRSA. There are limited data available against MRSA blood isolates (MRSABIs), vancomycin-intermediate strains (VISA), vancomycin-resistant strains (VRSA), daptomycin-non-susceptible strains (DNSSA) and linezolid-resistant Staphylococcus aureus (LRSA). METHODS: Antimicrobial activity of delafloxacin, levofloxacin, vancomycin, daptomycin and linezolid was determined against 110 MRSABIs, 15 VRSA, 35 VISA, 40 DNSSA and 6 LRSA. Microdilution testing using CAMHB was used to determine MIC according to CLSI guidelines. FDA breakpoints were used to determine delafloxacin susceptibility, and CLSI breakpoints were used for all other antibiotics. PCR testing for molecular markers was performed. RESULTS: Delafloxacin demonstrated activity against MRSABIs with an MIC90 of 1 mg/L and 68% susceptibility. Against the other groups the MIC90 and susceptibility were 1 mg/L and 40%, respectively, for VISA, 4 mg/L and 7% for VRSA and 1 mg/L and 38% for DNSSA. None of the LRSA isolates was susceptible to delafloxacin. Delafloxacin was active against 94% of MRSA blood isolates that were genotype SCC IVa. For MRSABIs with a levofloxacin MIC ≥8 mg/L (55/110), suggesting multiple mutations in the QRDR, delafloxacin MIC90 was 1 mg/L with a 36.4% susceptibility rate. CONCLUSIONS: Delafloxacin demonstrates superior activity to levofloxacin against recent MRSA blood isolates, VISA, VRSA and DNSSA, and demonstrates good activity against blood isolates most commonly found in the community.

TXA709, an FtsZ-Targeting Benzamide Prodrug with Improved Pharmacokinetics and Enhanced In Vivo Efficacy against Methicillin-Resistant Staphylococcus aureus.

The clinical development of FtsZ-targeting benzamide compounds like PC190723 has been limited by poor drug-like and pharmacokinetic properties. Development of prodrugs of PC190723 (e.g., TXY541) resulted in enhanced pharmaceutical properties, which, in turn, led to improved intravenous efficacy as well as the first demonstration of oral efficacy in vivo against both methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA). Despite being efficacious in vivo, TXY541 still suffered from suboptimal pharmacokinetics and the requirement of high efficacious doses. We describe here the design of a new prodrug (TXA709) in which the Cl group on the pyridyl ring has been replaced with a CF3 functionality that is resistant to metabolic attack. As a result of this enhanced metabolic stability, the product of the TXA709 prodrug (TXA707) is associated with improved pharmacokinetic properties (a 6.5-fold-longer half-life and a 3-fold-greater oral bioavailability) and superior in vivo antistaphylococcal efficacy relative to PC190723. We validate FtsZ as the antibacterial target of TXA707 and demonstrate that the compound retains potent bactericidal activity against S. aureus strains resistant to the current standard-of-care drugs vancomycin, daptomycin, and linezolid. These collective properties, coupled with minimal observed toxicity to mammalian cells, establish the prodrug TXA709 as an antistaphylococcal agent worthy of clinical development.

Ceftaroline-heteroresistant Staphylococcus aureus.

Heteroresistance refers to the presence, within a large population of antimicrobial-susceptible microorganisms, of subpopulations with lesser susceptibilities. Ceftaroline is a novel cephalosporin with activity against methicillin-resistant Staphylococcus aureus (MRSA). The aim of this study was to detect the prevalence of ceftaroline heteroresistance in vitro in a select group of S. aureus strains. There were 57 isolates selected for evaluation, 20 MRSA, 20 vancomycin-intermediate S. aureus (VISA), 7 daptomycin-nonsusceptible S. aureus (DNSSA), 6 linezolid-nonsusceptible S. aureus (LNSSA), and 4 heteroresistant VISA (hVISA) isolates. MICs and minimal bactericidal concentrations were determined using the broth microdilution method according to CLSI guidelines. All of the isolates were analyzed by pulsed-field gel electrophoresis. The staphylococcal cassette chromosome mec element (SCCmec) types were determined by a multiplex PCR. Population analysis profiles (PAPs) were performed to determine heteroresistance for all of the isolates using plates made by adding various amounts of ceftaroline to brain heart infusion agar. The frequencies of resistant subpopulations were 1 in 10(4) to 10(5) organisms. We determined that 12 of the 57 (21%) isolates tested were ceftaroline-heteroresistant S. aureus (CHSA). CHSA occurred among strains with reduced susceptibilities to vancomycin, daptomycin, and linezolid but occurred in none of the USA-300 isolates tested. Evaluation of the heteroresistant strains demonstrated that the phenotype was unstable. Further studies are needed to determine whether CHSA has a role in clinical failures and to determine the implications of our study findings.

Rhodomyrtone: a new anti-Staphylococcus aureus agent against resistant strains.

Background: Rhodomyrtone is a novel plant-derived antibiotic compound originally isolated from Rhodomyrtus tomentosa leaf extract. Objectives: To evaluate the activity of rhodomyrtone against a group of MRSA strains, including isolates with reduced susceptibility or resistance to vancomycin, daptomycin, linezolid and ceftaroline. Methods: Broth microdilution testing was used to determine the MICs and MBCs of rhodomyrtone, fosfomycin, vancomycin, daptomycin, linezolid and ceftaroline. Results: ROM had an MIC90 of 1 mg/L against 110 strains of MRSA from blood isolates as well as for all other isolates that were daptomycin resistant, VISA, VRSA or LRSA. The MBC90 were 4 mg/L across all groups tested. Among all S. aureus groups tested the ROM MBC did not exceed 8 mg/L. Conclusions: Rhodomyrtone demonstrated excellent activity against MRSA as well as isolates with resistance or reduced activity to other anti-MRSA drugs including vancomycin, daptomycin and linezolid. Rhodomyrtone may have potential clinical utility when treating patients with infections caused by MRSA including those with reduced susceptibility to first-line anti-MRSA antimicrobial agents.

In Vitro Activity of Retapamulin against Staphylococcus aureus Resistant to Various Antimicrobial Agents.

Retapamulin and six other antimicrobial agents were evaluated against 155 methicillin-resistant Staphylococcus aureus (MRSA) isolates, including strains resistant to vancomycin, linezolid, daptomycin, and mupirocin by microdilution tests. Time-kill assays were performed against representative MRSA, vancomycin-intermediate S. aureus (VISA), and vancomycin-resistant S. aureus (VRSA) isolates. Retapamulin and mupirocin demonstrated MIC90s of 0.12 µg/ml and 8 µg/ml, respectively, with resistance seen in 2.6% and 10% of isolates, respectively. Retapamulin maintained good activity against 94% (15/16) of mupirocin-resistant isolates.

In vitro susceptibilities and molecular analysis of vancomycin-intermediate and vancomycin-resistant Staphylococcus aureus isolates.

BACKGROUND: There is increasing frequency of vancomycin-intermediate and -resistant Staphylococcus aureus (VISA and VRSA) isolates identified in clinical practice. There are limited reports evaluating susceptibility patterns and molecular characteristics of these strains. METHODS: Laboratory analysis was performed on 13 VRSA and 33 VISA isolates, including susceptibility testing by broth microdilution, detection of Panton-Valentine leukocidin (PVL) genes, arginine catabolic mobile element (ACME), and staphylococcal cassette chromosome mec typing using polymerase chain reaction. Strain typing using pulsed-field gel electrophoresis (PFGE) was performed on VRSA isolates. RESULTS: Telavancin, linezolid, tigecycline, and minocycline were active against >90% of VISA isolates, while >90% of VRSA isolates were susceptible to ceftaroline, daptomycin, linezolid, minocyline, tigecycline, rifampin, and trimethoprim/sulfamethoxazole. There were no VISA or VRSA isolates that carried PVL genes or ACME, and most strains (69.8%) were staphylococcal cassette chromosome mec type II. VRSA isolates were predominantly related to USA100 (53.8%) and none were related to USA300 or USA400. CONCLUSIONS: A large number of available antimicrobial agents retain very good in vitro activity against VRSA and VISA isolates. The present isolates appear to be derived from healthcare-associated strains based on the absence of features associated with community-associated strains, and VRSA isolates are polyclonal by PFGE.

In vitro activities of LTX-109, a synthetic antimicrobial peptide, against methicillin-resistant, vancomycin-intermediate, vancomycin-resistant, daptomycin-nonsusceptible, and linezolid-nonsusceptible Staphylococcus aureus.

LTX-109 and eight other antimicrobial agents were evaluated against 155 methicillin-resistant Staphylococcus aureus (MRSA) isolates, including strains resistant to vancomycin and strains with decreased susceptibility to daptomycin and linezolid, by microdilution tests to determine MICs. Time-kill assays were performed against representative MRSA, vancomycin-intermediate S. aureus (VISA), and vancomycin-resistant S. aureus (VRSA) isolates. LTX-109 demonstrated a MIC range of 2 to 4 µg/ml and dose-dependent rapid bactericidal activity against S. aureus. This activity was not influenced by resistance to other antistaphylococcal agents.

A case of Gemella morbillorum native valve endocarditis and results of in vitro susceptibility testing.

We present a case of a 48 years old male with Gemella morbillorum native mitral valve endocarditis. Due to poor growth of the organism, antimicrobial susceptibility test (AST) could not be performed using the CLSI approved method. AST was determined using Etest© strips and the patient was successfully treated with mitral valve replacement and intravenous ceftriaxone.

In vitro activity of ceftaroline against community-associated methicillin-resistant, vancomycin-intermediate, vancomycin-resistant, and daptomycin-nonsusceptible Staphylococcus aureus isolates.

This study assessed the in vitro activities of ceftaroline and five comparator agents against a collection of Staphylococcus aureus isolates. Ceftaroline demonstrated potent activity against community-associated methicillin-resistant S. aureus (CA-MRSA) isolates and showed bactericidal activity against vancomycin-intermediate S. aureus (VISA), vancomycin-resistant S. aureus (VRSA), heteroresistant VISA (hVISA), and daptomycin-nonsusceptible S. aureus (DNSSA) isolates. Ceftaroline may represent a bactericidal treatment option for infections caused by these pathogens.

VISA-Daptomycin non-susceptible Staphylococcus aureus frequently demonstrate non-susceptibility to Telavancin.

Telavancin was evaluated against S. aureus isolates with reduced susceptibility to other antimicrobial agents using two broth microdilution methods and Etest® strips. The three methods provided comparable results. Differences in telavancin susceptibility versus non-susceptibility were noted mainly in the VISA-daptomycin non-susceptible group of isolates. In this group the percent susceptibility was 38% for the Etest® method and 50% and 54% for the 2 broth microdilution methods. All differences in susceptibility were within one 2-fold dilution.

Community-associated methicillin-resistant Staphylococcus aureus causing chronic pneumonia.

A young woman presented with pneumonia of a 3-month duration with predominantly nodular pulmonary infiltrates. Methicillin-resistant Staphylococcus aureus was identified in multiple cultures of sputum specimens. According to findings of pulsed-field gel electrophoresis, the isolate was identical to USA 300 and carried a type IV Staphylococcus cassette chromosome mec type IV gene and the genes for Panton-Valentine leukocidin.

Emergence of community-associated methicillin-resistant Staphylococcus aureus infection among patients with end-stage renal disease.

OBJECTIVE: To evaluate the frequency of infections due to community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) strains among our patients with end-stage renal disease. DESIGN: Prospective observational clinical and laboratory study of patients in 2005. Molecular features of isolates recovered from these patients were compared with those of isolates recovered in 2000 from patients with end-stage renal disease. SETTING: A 600-bed urban academic medical center. PATIENTS: Thirty-two patients with end-stage renal disease and MRSA infection at the time of hospitalization from 2005 were evaluated. For comparison, laboratory analysis was performed for 17 MRSA isolates recovered from patients with end-stage renal disease in 2000. RESULTS: The patients from 2005 were more likely than the patients from 2000 to have infection with strains that carried the staphylococcal cassette chromosome (SCC) mec type IV complex (50% vs 11.8%; relative risk, 4.25 [95% confidence interval, 1.17-25.98]; P = .012) and the Panton-Valentine leukocidin toxin genes (25% vs 0%; P = .038). Eight patients from 2005 were infected with a strain that is identical to MRSA clone USA300 in terms of molecular type and presence of SCCmec type IV and Panton-Valentine leukocidin genes. Among the patients from 2005, those infected with SCCmec type IV strains (ie, CA-MRSA strains) and those infected with SCCmec type II strains (ie, healthcare-associated MRSA [HA-MRSA] strains) were similar with respect to demographic characteristics, risk factors, and outcomes. CONCLUSIONS: We documented an increased proportion of infections with CA-MRSA strains, including clone USA300, among our population of patients undergoing dialysis. Patients infected with CA-MRSA strains and HA-MRSA strains were similar with respect to presenting illness and outcomes.

Clinical and laboratory features of community-associated methicillin-resistant Staphylococcus aureus: is it really new?

OBJECTIVE: To review the epidemiologic and molecular characteristics of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) in Detroit, Michigan, to assess the risk factors for infection and the response to therapy. DESIGN: Prospective clinical and laboratory study of 2003-2004 CA-MRSA isolates. Molecular features were compared with CA-MRSA isolates from 1980. SETTING: A 600-bed urban academic medical center. PATIENTS: Twenty-three patients with CA-MRSA infections from 2003-2004 were evaluated. In addition, laboratory analysis was performed on 13 CA-MRSA isolates from 1980. MAIN OUTCOME MEASURES: Laboratory analysis of isolates included antimicrobial susceptibility testing, pulsed-field genotyping, testing for Panton-Valentine leukocidin (PVL) genes, and staphylococcal cassette chromosome mec typing. RESULTS: Patients were predominantly young African American males and presented with skin and soft-tissue infections. All isolates were resistant to erythromycin and highly susceptible to other agents. Patients were generally treated successfully with combination incision and drainage and systemic antibiotics. Among the 23 isolates, 20 (87%) were the same strain. This strain carried the staphylococcal cassette chromosome mec type IV and PVL genes and is genetically identical to USA 300. Thirteen isolates of patients from our community who presented with CA-MRSA infections in 1980 represented a single clone that is unique compared with the 2003-2004 isolates. This strain carried staphylococcal cassette chromosome mec type IVA but did not carry the PVL genes. CONCLUSIONS: In our community, CA-MRSA is largely due to a single clone with a type IV mec gene and PVL gene. The type IV staphylococcal cassette chromosome mec type can be demonstrated in CA-MRSA isolates from a remote period, suggesting that earlier outbreaks were not related to healthcare exposure.

Antimicrobial activity and a comparison of published pharmacodynamics of gemifloxacin and eight fluoroquinolones against Streptococcus pneumoniae.

Gemifloxacin was evaluated for its in vitro activity and was compared with eight fluoroquinolones. Pharmacodynamic comparisons were made based on published pharmacokinetic information. Gemifloxacin demonstrated excellent in vitro activity (minimum inhibitory concentration necessary to inhibit 90% of the strains tested, MIC90 = 0.03 mg/L (range 0.0019-0.03 mg/L)) against 199 strains of Streptococcus pneumoniae. Its activity was not influenced by penicillin or ciprofloxacin non-susceptibility. Gemifloxacin demonstrated excellent pharmacodynamic parameters, with a Cmax/MIC90 of 67 (where Cmax is the peak serum level) and an AUC/MIC90 of 297 (where AUC is the area under the curve). Compared with the other eight fluoroquinolones tested, gemifloxacin demonstrated the best in vitro activity and Cmax/MIC90.

Broad-range bacterial polymerase chain reaction for early detection of bacterial meningitis.

The diagnosis of bacterial meningitis often depends on isolation of bacteria on culture, which may take 24-48 h. DNA amplification techniques could provide rapid diagnosis, which would guide the clinician in antimicrobial therapy decisions. This study determined the clinical utility of polymerase chain reaction (PCR) for the diagnosis of meningitis with use of a broad range of bacterial primers. Seventy-four cerebrospinal fluid specimens obtained from 70 patients were subjected to PCR with use of primers derived from conserved regions of the bacterial 16S RNA gene. The test characteristics for the broad-range bacterial PCR were as follows: sensitivity, 100%; specificity, 98.2%; positive predictive value, 94.4%; and negative predictive value, 100%. Broad-range bacterial PCR may be useful for excluding the diagnosis of meningitis, and the results may influence the decision to initiate or discontinue antimicrobial therapy.

Changing epidemiology of community-onset methicillin-resistant Staphylococcus aureus bacteremia.

OBJECTIVES: To review cases of community-onset Staphylococcus aureus bacteremia and to evaluate whether the risk factors and epidemiology of methicillin-resistant S. aureus (MRSA) bacteremia have changed from early reports. DESIGN: Retrospective case-comparison study of community-onset MRSA (n = 26) and methicillin-susceptible S. aureus (MSSA) (n = 26) bacteremias at our institution. SETTING: A 600-bed urban academic medical center. PATIENTS: Twenty-six patients with community-onset MRSA bacteremia were compared with 26 patients with community-onset MSSA bacteremia. Molecular analysis was performed on S. aureus isolates from the 26 MRSA cases as well as from 13 cases of community-onset S. aureus bacteremia from 1980 and 9 cases of nosocomial S. aureus bacteremia from 2001. RESULTS: The two groups were similar except that patients with MRSA bacteremia were more likely to have presented from a long-term-care facility (26.9% vs 4%; P = .05) and to have had multiple admissions within the preceding year (46% vs 15%; P = .03). Clamped homogeneous electric fields analysis of MRSA isolates from 1982 revealed predominantly that one clone was the epidemic strain, whereas there were 14 unique strains among current community-onset isolates. Among current nosocomial isolates, 3 patterns were identified, all of which were present in the community-onset cases. CONCLUSIONS: Previously described risk factors for MRSA acquisition may not be helpful in predicting disease due to the polyclonal spread of MRSA in the community. Unlike early outbreaks of MRSA in patients presenting from the community, current acquisition appears to be polyclonal and is usually related to contact with the healthcare system.

Rapid diagnosis of septic arthritis using 16S rDNA PCR: a comparison of 3 methods.

Few studies address the utility of molecular techniques for diagnosis of infection in synovial fluid (SF). We evaluated 3 different methods using 16S rDNA polymerase chain reaction (PCR) on 63 specimens for the diagnosis of joint infection. SF samples were classified as normal, inflammatory, or septic based on the patient's clinical and laboratory results. Samples were analyzed by conventional PCR using primers for the bacterial 16S rDNA gene and by real-time PCR utilizing 2 different sets of primers for the target gene 16S rDNA. PCR results were compared to culture results. All inflammatory and normal SF samples were culture negative. There was concordance with 10 of the 16 septic samples by 2 of the PCR methods. When comparing 3 methods for rapid detection of septic arthritis, real-time PCR using SYBR-Green I and conventional PCR demonstrated favorable test characteristics, but need further study.

In vitro activity of oritavancin against community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA), vancomycin-intermediate S. aureus (VISA), vancomycin-resistant S. aureus (VRSA) and daptomycin-non-susceptible S. aureus (DNSSA).

Isolates of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA), vancomycin-intermediate S. aureus (VISA), vancomycin-resistant S. aureus (VRSA) and daptomycin non-susceptible S. aureus (DNSSA) are increasing in frequency and new antistaphylococcal therapies are needed. Microdilution testing using Mueller-Hinton broth was used to determine the minimal inhibitory concentrations (MICs) of oritavancin and nine additional antimicrobial agents against 92 CA-MRSA, 23 VISA, 7 DNSSA and 10 VRSA isolates. Minimal bactericidal concentrations were also determined. Pulsed-field gel electrophoresis (PFGE) was performed. Staphylococcal cassette chromosome mec (SCCmec) typing as well as assays for Panton-Valentine leukocidin (PVL) and arginine catabolic mobile element (ACME) genes were performed. Oritavancin was more bactericidal than any of the other comparators against CA-MRSA and demonstrated excellent activity against VRSA and VISA.

Prevalence of colonization with community-associated methicillin-resistant Staphylococcus aureus among end-stage renal disease patients and healthcare workers.

OBJECTIVE: To evaluate the prevalence, epidemiologic features, and molecular characteristics of colonization with community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) among hospitalized dialysis patients and their healthcare workers (HCWs). DESIGN: Prospective observational clinical and laboratory study of nasal colonization. SETTING: A 600-bed urban academic medical center. SUBJECTS: One hundred twenty hospitalized dialysis inpatients and 100 HCWs. RESULTS: Of 120 patients, 40 (33%) were colonized with S. aureus; 26 (65%) of these 40 were colonized with MRSA. Among the 26 MRSA isolates, 10 (38.5%) carried staphylococcal cassette chromosome (SCC) mec type IV (ie, CA-MRSA), and 7 of these 10 carried the genes for the Panton-Valentine leukocidin (PVL) toxin. Patients colonized with healthcare-associated MRSA strains and those colonized with CA-MRSA strains were similar, except for a higher frequency of a history of congestive heart failure among those with healthcare-associated MRSA strains (P=.014). Among 10 patients who presented with or developed an S. aureus infection while hospitalized, 8 were colonized with S. aureus, 7 with MRSA, and 3 with SCCmec type IV strains. Among 100 HCWs, 31 were colonized with S. aureus, including 6 with MRSA; 2 of the MRSA isolates belonged to CA-MRSA strains, and soft-tissue infections were reported in one of the HCWs and in the family member of the other HCW colonized with these strains. CONCLUSIONS: There is a high rate of colonization with MRSA and CA-MRSA among hospitalized dialysis patients and their HCWs. As other studies have found, it appears that individuals are being colonized with both CA-MRSA strains and healthcare-associated MRSA strains.