Bloodstream infections due to Candida species: SENTRY antimicrobial surveillance program in North America and Latin America, 1997-1998

An international program of surveillance of bloodstream infections (BSI) in the United States, Canada, and Latin America detected 306 episodes of candidemia in 34 medical centers (22 in the United States, 6 in Canada, and 6 in Latin America) in 1997 and 328 episodes in 34 medical centers (22 in the United States, 5 in Canada, and 7 in Latin America) in 1998. Of the 634 BSI, 54.3% were due to Candida albicans, 16.4% were due to C. glabrata, 14.9% were due to C. parapsilosis, 8.2% were due to C. tropicalis, 1.6% were due to C. krusei, and 4.6% were due to other Candida spp. The percentage of BSI due to C. albicans decreased very slightly in the United States between 1997 and 1998 (56.2 to 54.4%; P = 0.68) and increased in both Canada (52.6 to 70.1%; P = 0.05) and Latin America (40.5 to 44. 6%; P = 0.67). C. glabrata was the second most common species observed overall, and the percentage of BSI due to C. glabrata increased in all three geographic areas between 1997 and 1998. C. parapsilosis was the third most prevalent BSI isolate in both Canada and Latin America, accounting for 7.0 and 18.5% of BSI, respectively. Resistance to fluconazole (MIC, >/=64 microgram/ml) and itraconazole (MIC, >/=1.0 microgram/ml) was observed infrequently in both 1997 (2.3 and 8.5%, respectively) and 1998 (1.5 and 7.6%, respectively). Among the different species of Candida, resistance to fluconazole and itraconazole was observed in C. glabrata and C. krusei, whereas isolates of C. albicans, C. parapsilosis, and C. tropicalis were all highly susceptible to both fluconazole (98.9 to 100% susceptible) and itraconazole (96.4 to 100% susceptible). Isolates from Canada and Latin America were generally more susceptible to both triazoles than U.S. isolates were. Continued surveillance appears necessary to detect these important changes.     

Emergence in Italy of a Neisseria meningitidis clone with decreased susceptibility to penicillin

A rise in invasive diseases due to Neisseria meningitidis C:2b:P1.5 with decreased penicillin susceptibility occurred in Italy during the last 2 years. Real-time PCR identified the Peni phenotype, and the penA sequence revealed the mosaicism of the gene. Molecular analyses assigned the isolates to a single emergent clone of the hypervirulent A4 cluster.     

Antimicrobial resistance and molecular epidemiology of vancomycin-resistant enterococci from North America and Europe: a report from the SENTRY antimicrobial surveillance program

Increases in prevalence of vancomycin-resistant enterococci (VRE) have been documented globally since its emergence in the 1980s. A SENTRY Antimicrobial Surveillance Program (2003) objective monitored VRE isolates with respect to antimicrobial susceptibility trends, geographic resistance variability, and clonal dissemination. In 2003, VRE isolates from North America (United States and Canada, n = 839, 26 sites) and Europe (n = 56, 10 sites) were susceptibility tested using Clinical and Laboratory Standards Institute (CLSI) reference methodologies. Based on resistance profiles, 155 isolates displayed similar multidrug-resistant (MDR) profiles and were temporally related; these were subsequently submitted for typing by pulsed-field gel electrophoresis (PFGE). Most of the submitted isolates were Enterococcus faecium (91.0%) and Enterococcus faecalis (7.8%). Among VRE, the VanA phenotype was more prevalent in North America (76%) than Europe (40%), and all isolates had elevated resistance rates to other antimicrobial classes including the following: 1) chloramphenicol resistance among E. faecalis being greater in North America than in Europe (28.6% versus 7.1%, respectively) but reversed among E. faecium (0.5% and 15.0%, the latter due to clonal occurrences); 2) ciprofloxacin resistance in North America >99% for both species and in Europe varying from 85.7% to 87.5%; 3) rare occurrences of linezolid resistance in North America (0.8% to 1.8%) due to G2576U ribosomal mutation; 4) higher quinupristin/dalfopristin resistance observed among European E. faecium strains (10.0% versus 0.6%); and 5) higher rifampin resistance rates among European E. faecalis (21.4% versus 5.4%). Thirty-five MDR epidemic clusters were identified by PFGE in 21 North American and 2 European medical centers including the following: 1) VanA (20 sites, 27 clonal occurrences) and VanB (1 site, 2 clonal occurrences); 2) elevated quinupristin/dalfopristin MIC results (not vatD/E, 3 sites); and 3) chloramphenicol resistance (chloramphenicol acetyltransferase-positive strains, 3 sites). The esp gene, part of the putative E. faecium pathogenicity island and a marker for the clonal complex-17 lineage, was detected in 76% of vancomycin-resistant E. faecium. Clonal spread appears to be a dominant factor of MDR VRE dissemination on both continents, and further monitoring is critical to assist in the control of these resistant pathogens.     

Fluoroquinolone-resistant Moraxella catarrhalis in a patient with pneumonia: report from the SENTRY antimicrobial surveillance program (1998)

Fluoroquinolone resistance in Moraxella catarrhalis isolates has been quite rare. This report presents a case history of a 22-year-old man with compromised immune status and severe pneumonia caused by M. catarrhalis. The organism was markedly resistant (MICs, 1.5–>32 μg/mL) to several marketed fluoroquinolones including the agent (levofloxacin) used for concurrent and prior therapy. The emergence of this problematic strain seems related to chronic exposure of the patient to compounds in the class and poor patient compliance to prescribed medications. The strain was not clonally related to other M. catarrhalis strains isolated in the same hospital during early 1998. This second documented case of a fluoroquinolone-resistant M. catarrhalis clinical isolate presents a warning that resistances can emerge in at-risk patients, and that surveillance systems (SENTRY) will be necessary to monitor for unusual organisms and spread of resistance phenotypes among commonly isolated respiratory tract pathogens.     

Five-year analysis of Haemophilus influenzae isolates with reduced susceptibility to fluoroquinolones: prevalence results from the SENTRY antimicrobial surveillance program

The appearance of resistance or reduced susceptibility to fluoroquinolones among Hemophilus influenzae has been documented for nearly a decade. Over this time, the use of fluoroquinolones for the treatment of respiratory infections including commonly isolated bacterial causes of community-acquired infections has markedly increased. The documentation of resistance to fluoroquinolones among Streptococcus pneumoniae and H. influenzae has also become more prevalent as measured by peer-reviewed publications. During 1997-2001, a total of 11,355 H. influenzae isolates were tested by reference broth microdilution methods from strains collected by the SENTRY Antimicrobial Surveillance Program (American and European medical centers). Strains with reduced susceptibility to fluoroquinolones (RSF) were detected during all five study years at an overall rate of 0.15%. Among the tested compounds, sitafloxacin (MIC(50,) 0.03 microg/ml) was the most potent agent against the RSF strains, followed by gemifloxacin (0.12 microg/ml) > garenoxacin = grepafloxacin = levofloxacin = moxifloxacin = trovafloxacin (0.5 microg/ml) > ciprofloxacin = sparfloxacin (1 microg/ml). Gene sequencing of the quinolone resistance determining region and epidemiologic typing of 30 RSF isolates showed diverse mutational events in gyr A and par C and multiple pulsed-field gel electrophoresis (PFGE) patterns among strains that was not consistent with clonal dissemination. Continued surveillance by global or national networks should continue to monitor for H. influenzae isolates that are refractory to fluoroquinolone therapy.     

Frequency of occurrence and antimicrobial susceptibility patterns for pathogens isolated from latin american patients with a diagnosis of pneumonia: results from the SENTRY antimicrobial surveillance program (1998)

The correct empiric choice of antimicrobial therapy in the treatment of pneumonia in hospitalized patients has established itself as a major therapeutic challenge to clinicians. Selection of an inappropriate antimicrobial agent could lead to increased rates of mortality and morbidity. Characteristics of pathogens responsible for this infection such as species prevalence, overall antimicrobial resistance rates, and mechanisms of detected resistance could serve as an invaluable resource to clinicians in making such therapeutic selections. This report addresses the aforementioned problems/needs by analysis of 712 strains isolated from the lower respiratory tract of patients hospitalized with a diagnosis of pneumonia in 10 Latin American medical centers in the SENTRY Antimicrobial Surveillance Program (1998). The four most frequently isolated pathogens (no/% of total) were: Pseudomonas aeruginosa (191/26.8%), Staphylococcus aureus (171/24.0%), Klebsiella spp. (86/12.1%), and Acinetobacter spp. (75/10.5%); representing nearly 75.0% of all isolates. More than 40 antimicrobial agents (23 reported) were tested against these isolates by reference broth microdilution methodology, and susceptibility profiles were established. The nonfermentative Gram-negative bacteria (P. aeruginosa and Acinetobacter spp.) exhibited high levels of resistance to the agents tested. Amikacin (77.5% susceptible) was the most active drug tested against P. aeruginosa 50.0% against the Acinetobacter spp. isolates. Based on published interpretive criteria, over 22.0% of the Klebsiella spp. and 12.5% of the Escherichia coli were classified as extended spectrum beta-lactamase (ESBL) producers. Of the cephalosporin class compounds tested against the Klebsiella spp. and E. coli isolates, cefepime demonstrated the highest rates of susceptibility (84.9% and 91.7%, respectively). This compound also fared well against the Enterobacter spp. isolates, inhibiting 88.2% of the isolates tested, many of which were resistant to ceftazidime and ceftriaxone. Resistance to oxacillin among the S. aureus isolates was nearly 50. 0%, with vancomycin, teicoplanin, and the streptogramin combination quinupristin/dalfopristin inhibiting all isolates. Several clusters of multiply resistant organisms were also observed, and further characterization by ribotyping and pulsed-field gel electrophoresis established possible patient-to-patient spread. The results of this study indicate that rates of resistance among respiratory tract pathogens continue to rise in Latin America, with specific concerns for the high prevalence of nonfermentative Gram-negative bacteria isolated, oxacillin resistance rates in S. aureus, and the epidemic dissemination of multiply-resistant strains in several medical centers. International surveillance programs (SENTRY) should assist in the control of escalating antimicrobial resistance in this geographic area.     

Polymorphism of Neisseria meningitidis penA gene associated with reduced susceptibility to penicillin

We studied polymorphism of penA (which encodes penicillin-binding protein 2) in 13 strains of Neisseria meningitidis susceptible to penicillin (pen(S)) and 12 strains with reduced susceptibility to penicillin (pen(I)). These strains differed in geographical origin. Serological and genetic typing showed that they were highly diverse and belonged to several genetic lineages. Restriction analysis and DNA sequencing of penA showed that all pen(S) strains had the same penA allele regardless of genetic group, whereas pen(I) strains harboured various penA alleles. Transformation with amplicons of penA and genomic DNA from several pen(I) strains conferred the pen(I) phenotype on a pen(S) strain. Thus, reduction in susceptibility to penicillin is directly related to changes in penA and analysis of penA polymorphisms could be used as a reliable tool for characterizing meningococcal strains in terms of their susceptibility to penicillin.     

Urinary tract infection trends in Latin American hospitals: report from the SENTRY antimicrobial surveillance program (1997-2000)

Urinary tract infections (UTI) are one of the most common infectious diseases diagnosed in outpatients as well as in hospitalized patients. The objective of this study was to report the frequency of occurrence and antimicrobial susceptibility of uropathogens collected in Latin America between 1997 to 2000 through the SENTRY Antimicrobial Surveillance Program. Antimicrobial susceptibility testing was performed and results interpreted using reference broth microdilution methods. In the 4 year period, a total of 1961 urine isolates from hospitalized patients were included. The patients' mean age was 51.3 years and most of the infections occurred among women (65.6%). Esherichia coli was the most frequent pathogen isolated followed by Klebsiella spp., Pseudomonas aeruginosa, and Proteus mirabilis. Among the E. coli isolates, piperacillin/tazobactam, aztreonam, extended-spectrum cephalosporins, carbapenems and amikacin constitute reasonable therapeutic options for treatment of serious UTI in Latin America (91.0-100.0% susceptible). High resistance rates to fluoroquinolones (17.5-18.9%) and trimethoprim/sulfamethoxazole (>45.0%) were observed among the E. coli. In contrast, nitrofurantoin displayed susceptibility rate of > 87.0%. Against Klebsiella spp. infections, the only effective therapeutic option would be the carbapenems due to the high number of isolates (>30.0%) producing extended-spectrum beta-lactamases (ESBL). Even the new fluoroquinolones showed limited activity against Klebsiella spp. (72.1-88.6% susceptible) and the P. aeruginosa isolates showed high resistance rates to most antimicrobial agents tested. The results of this survey endorse the importance of Enterobacteriaceae as cause of UTI in Latin America. Our results also demonstrate that the uropathogens isolated in the Latin American medical centers exhibit high resistance to various classes of antimicrobial agents. Carbapenem-resistant P. aeruginosa, ciprofloxacin-resistant E. coli, ESBL-producing K. pneumoniae constitute serious problem in this geographic region.     

Activity and spectrum of 22 antimicrobial agents tested against urinary tract infection pathogens in hospitalized patients in Latin America: report from the second year of the SENTRY antimicrobial surveillance program (1998)

The potency and spectrum of various antimicrobial agents tested against 434 bacterial isolates causing urinary tract infection (UTI) in hospitalized patients in Latin America were evaluated. The genotypes of the extended-spectrum beta-lactamase-producing and selected multi-resistant isolates were also evaluated by molecular typing techniques. Escherichia coli (60.4%) was the most common aetiological agent causing UTI, followed by Klebsiella spp. (11.2%) and Pseudomonas aeruginosa (8.3%). In contrast, Enterococcus spp. isolates caused only 2.3% of UTIs. Fewer than 50% of E. coli isolates were susceptible to broad-spectrum penicillins. The resistance rates to ciprofloxacin and the new quinolones were also high among these isolates. The molecular characterization of ciprofloxacin-resistant E. coli showed that most of them have a double mutation in the gyrA gene associated with a single mutation in the parC gene. The Klebsiella pneumoniae isolates studied demonstrated high resistance rates to beta-lactam drugs, including broad-spectrum cephalosporins. The carbapenems were the compounds with the highest susceptibility rate among these isolates (100.0% susceptible) followed by cefepime (91.7% susceptible). Meropenem, imipenem and cefepime were also the most active drugs against Enterobacter spp. Among P. aeruginosa isolates, meropenem (MIC(50), 2 mg/L) was the most active compound, followed by imipenem (MIC(50), 4 mg/L), cefepime (MIC(50), 8 mg/L) and ceftazidime (MIC(50), 16 mg/L). The results presented in this report confirm that bacterial resistance continues to be a great problem in Latin American medical institutions.     

Activity of gatifloxacin tested against isolates from pediatric patients: report from the SENTRY Antimicrobial Surveillance Program (North America, 1998-2003)

The SENTRY Antimicrobial Surveillance Program has monitored the activity of antimicrobial agents worldwide since 1997. The increasing number of clinical failures with established anti-infectives (penicillins, other beta-lactams, macrolides) among pediatric patients has stressed the importance for alternative therapeutic options. Ciprofloxacin has been recently approved for expanded use as treatment of complicated urinary tract infections in children, and gatifloxacin has been used successfully in clinical trials in selected children with severe or refractory otitis media. We evaluated the activity of gatifloxacin against strains isolated from children < 7 years of age and compared this to the general patient population (all ages included) using the SENTRY Program database. A total of 59826 North American isolates were collected, of which 4641 were from children (< 7 years old); all isolates were tested using reference broth microdilution methods. In contrast to the general population (GP), gatifloxacin resistance rates were very low among isolates from this younger patient group. Gatifloxacin susceptibility rates were > 84% for all pathogens evaluated in younger patients. All Streptococcus pneumoniae strains from children < 7 years old were susceptible to gatifloxacin, and susceptibility among the Enterobacteriaceae species was > 98%. The greatest difference in susceptibility rates between the younger children and the GP was observed among nonfermentative gram-negative bacilli (95.0-100% versus 64.8-83.7%, respectively) and Enterococcus faecalis (94.7% versus 58.4%). Gatifloxacin susceptibilities of Pseudomonas aeruginosa and Acinetobacter spp. isolates from the pediatric population were > or = 95% (> 97% for ciprofloxacin) compared to the GP at only 64.8-69.1%. In conclusion, gatifloxacin remains very active against bacterial isolates from children < 7 years, indicative of the limited exposure of this population to fluoroquinolones. Continued resistance surveillance will be necessary to monitor the activity of the fluoroquinolone class as they are introduced for specific clinical indications into the pediatric age groups, especially if re-studied against S. pneumoniae (refractory otitis media) and P. aeruginosa (cystic fibrosis associated pneumonia).     

Target gene sequencing to characterize the penicillin G susceptibility of Neisseria meningitidis

Clinical isolates of Neisseria meningitidis with reduced susceptibility to penicillin G (intermediate isolates, Pen(I)) harbor alterations in the penA gene encoding the penicillin binding protein 2 (PBP2). A 402-bp DNA fragment in the 3' half of penA was sequenced from a collection of 1,670 meningococcal clinical isolates from 22 countries that spanned 60 years. Phenotyping, genotyping, and the determination of MICs of penicillin G were also performed. A total of 139 different penA alleles were detected with 38 alleles that were highly related, clustered together in maximum-likelihood analysis and corresponded to the penicillin G-susceptible isolates. The remaining 101 penA alleles were highly diverse, corresponded to different genotypes or phenotypes, and accounted for 38% of isolates, but no clonal expansion was detected. Analysis of the altered alleles that were represented by at least five isolates showed high correlation with the Pen(I) phenotype. The deduced amino acid sequence of the corresponding PBP2 comprised five amino acid residues that were always altered. This correlation was not complete for rare alleles, suggesting that other mechanisms may also be involved in conferring reduced susceptibility to penicillin. Evidence of mosaic structures through events of interspecies recombination was also detected in altered alleles. A new website was created based on the data from this work (http://neisseria.org/nm/typing/penA). These data argue for the use of penA sequencing to identify isolates with reduced susceptibility to penicillin G and as a tool to improve typing of meningococcal isolates, as well as to analyze DNA exchange among Neisseria species.     

High prevalence of oxacillin-resistant Staphylococcus aureus isolates from hospitalized patients in Asia-Pacific and South Africa: results from SENTRY antimicrobial surveillance program, 1998-1999

As part of the SENTRY antimicrobial surveillance program, we examined the prevalence rates, types, and antibiograms of oxacillin-resistant Staphylococcus aureus from hospitalized patients from 17 institutions in eight countries in Asia-Pacific and South Africa (APAC). From April 1998 to December 1999, a total of 1,711 isolates of S. aureus (814 from blood, 392 from the respiratory tract, 467 from skin and skin structures, and 38 from urine) were collected from hospitalized patients within the APAC region. Multidrug-resistant oxacillin-resistant S. aureus (MORSA) isolates, defined as strains with three or more resistances to drug classes other than beta-lactams, were the most common type of oxacillin-resistant S. aureus (ORSA). They were the most frequently identified pathogen in wound infections and were common in bloodstream and lower respiratory tract infections. In all contributing institutions combined, more than 45% (range, 4 to 74%) of S. aureus isolates were oxacillin resistant, and in six institutions, this rate exceeded 60%. MORSA accounted for 91.2% of all oxacillin-resistant isolates. Distinct resistance patterns predominated at various sites within the APAC region, suggesting the local evolution of resistant clones. Non-multidrug-resistant strains were frequent in one part of Australia. No vancomycin-intermediate strains were detected, and no strains were resistant to linezolid or quinupristin-dalfopristin. MORSA strains are a very common cause of infection in hospitalized patients in the APAC region.     

Molecular characterization of SPM-1, a novel metallo-beta-lactamase isolated in Latin America: report from the SENTRY antimicrobial surveillance programme

The gene encoding the metallo-beta-lactamase SPM-1 was cloned from a genomic library of Pseudomonas aeruginosa strain 48-1997 A. The insert carrying spm-1 possessed a GC content of 47%, indicating that it is of non-Pseudomonas origin. Upstream of spm-1 there is a small open reading frame (ORF), which is homologous to the LysR family of proteins (69% identity to the LysR protein from Salmonella enterica serovar Typhimurium). Downstream of spm-1 there is the start of an ORF, the product of which shows close homology with the GroEL-type proteins from Xanthomonas campestris. No transmissible element could be identified upstream or downstream of spm-1. The spm-1 gene is carried on a plasmid that can transform both Escherichia coli and P. aeruginosa to ceftazidime resistance. SPM-1 contains the classic metallo-beta-lactamase zinc-binding motif HXHXD and shows the highest identity (35.5%) to IMP-1. SPM-1 is a distinctly different metallo-beta-lactamase from VIM and IMP and, accordingly, represents a new subfamily of mobile metallo-beta-lactamases. The predicted molecular weight of the protein was 27 515 Da, significantly higher than that of IMP (25 041 Da) or VIM (25 322 Da). SPM-1 possesses a unique loop of 23 residues that accounts for the higher molecular mass.     

Detection of resistance to rifampicin and decreased susceptibility to penicillin in Neisseria meningitidis by real-time multiplex polymerase chain reaction assay

In this study, we designed primers and probes for the rpoB gene of Neisseria meningitidis to detect rifampicin-resistant strains in a combined use with primers and probes previously described for penicillin intermediate isolates. The assay was set up in the Light Cycler instrument using the Fluorescence Resonance Energy Transfer platform. The method was applied to N. meningitidis strains and to culture-negative cerebrospinal fluids (CSFs) from patients with meningococcal invasive disease. A spiked CSF was used to determine the sensitivity of the assay.     

Fluoroquinolone-resistant Haemophilus influenzae: frequency of occurrence and analysis of confirmed strains in the SENTRY antimicrobial surveillance program (North and Latin America)

The incidence of fluoroquinolone-resistant (FQR) Haemophilus influenzae and Moraxella catarrhalis isolated from clinical specimens remains very rare, and the identification of such strains has been previously limited to case reports from diverse geographic locations. During the 1997 through 1998 SENTRY Antimicrobial Surveillance Program, four FQR-H. influenzae (0.13% of all strains) and one FQR-M. catarrhalis strains were identified and confirmed as having elevated MICs to > or =5 FQ class drugs. Among H. influenzae strains, MICs to marketed FQs were > or =0.12 microg/ml with ciprofloxacin MIC results > or =8-fold higher than wild type susceptible strains. The FQR-H. influenzae isolates were then compared with two previously reported strains that were determined to be identical using ribotyping and other molecular methods. In contrast, the SENTRY isolates were all genetically distinct and had mutations in parC and/or gyrA. Isolates having the lowest MIC elevations had a single mutation in gyrA, while isolates with higher MICs had at least one mutation in both studied genes. In general, the single gyr A mutations involved the same position but differed in the amino acid substitution (Ser84Leu or Phe or Ala). The isolates reported outside the SENTRY Program (controls) had an unusual mutation in par C (Gly82Asp) and two mutations in gyr A; producing the highest recorded FQR MICs. The FQR-M. catarrhalis strain was discovered in late 1997 and has been reported before. Although detection of these FQR isolates remains at <1% of all contemporary H. influenzae and M. catarrhalis isolates, surveillance programs will be an important detection method to determine the extent of emerging novel resistance patterns among clinically prevalent fastidious pathogens.     

Comparative activity and spectrum of broad-spectrum beta-lactams (cefepime, ceftazidime, ceftriaxone, piperacillin/tazobactam) tested against 12,295 staphylococci and streptococci: report from the SENTRY antimicrobial surveillance program (North America: 2001-2002)

A contemporary collection of 12,295 North American isolates (2001-2002) consisting of Staphylococcus aureus (50%), coagulase-negative staphylococci (12%), Streptococcus pneumoniae (24%), beta-hemolytic streptococci (12%), and viridans-group streptococci (2%) were tested against broad-spectrum beta-lactams (cefepime, ceftazidime, ceftriaxone, imipenem, piperacillin/tazobactam) and comparator agents using a reference broth microdilution method to determine their continued effectiveness for empiric antimicrobial therapy. All isolates were very susceptible to vancomycin, linezolid and quinupristin/dalfopristin (>98%). Oxacillin-susceptible staphylococci were also highly susceptible to the tested beta-lactams (>98%) with the exception of ceftazidime (93%). beta-hemolytic streptococci were exquisitely susceptible (>99%) to penicillin and all other agents except for clindamycin (94%) and erythromycin (81%). Viridans group streptococci were routinely less susceptible than were other streptococci. S. pneumoniae remained susceptible to most agents (>91%) with the exceptions of erythromycin (74%) and penicillin (69%). Among beta-lactams tested against S. pneumoniae, ceftriaxone and cefepime continued to be very active against penicillin-susceptible (>99%) and intermediate (>98%) strains, but less active (80% and 82%, respectively) against penicillin-resistant isolates. These findings confirm that the newer cephalosporins (cefepime and ceftriaxone) among broad-spectrum beta-lactam agents have a spectrum of activity that remains comprehensive for the commonly isolated Gram-positive pathogens.