Etest MICs Research Articles

Characterization of heterogeneous vancomycin-intermediate resistance, MIC and accessory gene regulator (agr) dysfunction among clinical bloodstream isolates of staphyloccocus aureus

BackgroundThe development of hVISA has been associated with vancomycin clinical failures and is commonly misidentified in clinical microbiology laboratories. Therefore, the objectives of this present study was to improve the reliability of methodologies and criteria for identifying hVISA, evaluate the prevalence of hVISA among clinical bloodstream isolates of S. aureus and determine if there exists a relationship between accessory gene regulator (agr) dysfunction and the hVISA phenotype.MethodsThe presence of hVISA in 220 clinical S. aureus isolates (121 MSSA, 99 MRSA) from bloodstream infections was examined by CLSI broth microdilution, Macro & Standard Etest. Isolates which were classified as hVISA by Macro Etest, were additionally evaluated using a modified PAP-AUC method using a modified starting inoculum of 1010 CFU/mL, and growth on brain heart infusion agar with 4 mg/L vancomycin (BHIV4) at 108 and 1010 CFU/mL, and agr function was assessed by delta-hemolysin production.ResultsBroth microdilution MIC50/90 of S.aureus and hVISA was 1.0/2.0 and 1.5/2.0 mg/L (p= 0.02), respectively. Macro Etest identified 12 (5.5%) hVISA isolates; higher among MRSA (9.1%) versus MSSA (2.5%) (p = 0.03). The mean modified PAP-AUC ratios (> 0.8) of 7 MRSA strains and 3 MSSA strains were significantly different (p = 0.001). 58% of hVISA strains were found to be agr dysfunctional when 21% of MRSA strains were agr dysfunctional. hVISA was detected among S. aureus bloodstream isolates, which were classified as susceptible among clinical microbiology laboratories.ConclusionsEvaluating the correlation between Etest MICs and modified PAP-AUC ratio values will add further improvement of discriminating hVISA, and agr dysfunction may be predictive of strains which display a greater predilection to display the hVISA phenotype.

Cryptic species and azole resistance in the Aspergillus niger complex.

Aspergillus niger is a common clinical isolate. Multiple species comprise the Aspergillus section Nigri and are separable using sequence data. The antifungal susceptibility of these cryptic species is not known. We determined the azole MICs of 50 black aspergilli, 45 from clinical specimens, using modified EUCAST (mEUCAST) and Etest methods. Phylogenetic trees were prepared using the internal transcribed spacer, beta-tubulin, and calmodulin sequences to identify strains to species level and the results were compared with those obtained with cyp51A sequences. We attempted to correlate cyp51A mutations with azole resistance. Etest MICs were significantly different from mEUCAST MICs (P < 0.001), with geometric means of 0.77 and 2.79 mg/liter, respectively. Twenty-six of 50 (52%) isolates were itraconazole resistant by mEUCAST (MICs > 8 mg/liter), with limited cross-resistance to other azoles. Using combined beta-tubulin/calmodulin sequences, the 45 clinical isolates grouped into 5 clades, A. awamori (55.6%), A. tubingensis (17.8%), A. niger (13.3%), A. acidus (6.7%), and an unknown group (6.7%), none of which were morphologically distinguishable. Itraconazole resistance was found in 36% of the isolates in the A. awamori group, 90% of the A. tubingensis group, 33% of the A. niger group, 100% of the A. acidus group, and 67% of the unknown group. These data suggest that cyp51A mutations in section Nigri may not play as important a role in azole resistance as in A. fumigatus, although some mutations (G427S, K97T) warrant further study. Numerous cryptic species are found in clinical isolates of the Aspergillus section Nigri and are best reported as "A. niger complex" by clinical laboratories. Itraconazole resistance was common in this data set, but azole cross-resistance was unusual. The mechanism of resistance remains obscure.

Relevance of vancomycin-intermediate susceptibility and heteroresistance in methicillin-resistant Staphylococcus aureus bacteraemia

To assess the relevance of vancomycin-intermediate susceptibility (VISA) and heteroresistance (hVISA) in methicillin-resistant Staphylococcus aureus (MRSA) bacteraemia. We determined vancomycin MICs for 371 saved MRSA blood isolates (2002-03; 2005-06) by Etest and broth microdilution (BMD), screened for hVISA (Etest methods), determined the population analysis profile (PAP)/AUC for isolates with suspected reduced susceptibility (MICs >2 mg/L and/or hVISA-screen-positive versus Mu3 (hVISA control), and stratified patient characteristics and outcome according to susceptibility phenotype: VISA (PAP/AUC >1.3), hVISA (PAP/AUC 0.9-1.3), and susceptible (S-MRSA; PAP/AUC <0.9). PAP/AUC revealed 6 (1.6%) VISA and 30 (8.1%) hVISA phenotypes. The Etest MIC was above the susceptibility cut-off (2 mg/L) for all VISA isolates, whereas the BMD MIC was within the susceptibility range in two (33.3%) instances. Eight hVISA isolates (26.7%) with MICs of 2 mg/L were hVISA-screen negative. SCCmec typing revealed SCCmec II in 100% of VISA, 86.7% of hVISA and 75.5% of S-MRSA isolates (P = 0.04). Prior vancomycin use was documented in 100% of VISA, 73.3% of hVISA and 52.2% of S-MRSA cases (P = 0.002). Outcome (compared in 243 vancomycin-treated patients with MICs of 2 mg/L) revealed longer time to clearance in VISA cases [12.1 ± 13.1 days versus 3.3 ± 3.9 (hVISA) and 3.7 ± 5.1 (S-MRSA); P = 0.001], more frequent endocarditis [33.3% versus 9.1% (hVISA; P = 0.1) and 4.2% (S-MRSA; P = 0.001)] and attributable mortality [33.3% versus 9.1% (hVISA; P = 0.1) and 8.4% (S-MRSA); P = 0.08]. No adverse outcome was documented with hVISA phenotype, whereas VISA contributed to vancomycin treatment failure. VISA and hVISA appear to emerge in SCCmec II isolates among vancomycin-exposed patients and are better detected by Etest.

Comparison of micafungin MICs as determined by the Clinical and Laboratory Standards Institute broth microdilution method (M27-A3 document) and Etest for Candida spp. isolates

Micafungin Etest and Clinical and Laboratory Standards Institute (CLSI) MICs were compared for 337 Candida spp. isolates. The performance of Etest for testing the susceptibilities of Candida spp. to micafungin was evaluated by the assessment of both categorical (CA) and essential (EA) agreements. The CA was evaluated 2 ways: (i) by the ability of Etest to separate resistant (nontreatable) from susceptible (treatable) isolates by using the newly adjusted species-specific micafungin clinical breakpoints (CBPs) that are available for most of the common species tested and (ii) by the ability to separate wild type (WT) from non-WT isolates or those harboring FKS mutations (with reduced echinocandin susceptibility) by using micafungin epidemiologic cutoff values (ECVs). Etest and CLSI MICs were in EA when the MICs were within 2 log2 dilutions. Based on agreement percentages, our data indicated that Etest is suitable to test micafungin for most of the Candida species evaluated (overall EA 94.7%; overall CA according to CBPs 97.2% and according to ECVs 97.3%). However, the number of resistant isolates was small, so further evaluations are needed with a higher number of such isolates including more resistant or those with known mechanisms of resistance (non-WT).

Determination of Vancomycin and Daptomycin MICs by Different Testing Methods for Methicillin-Resistant Staphylococcus aureus

Vancomycin and daptomycin MICs from 161 isolates of methicillin-resistant Staphylococcus aureus (MRSA) were compared using commercial and in-house broth microdilution, Etest, and common automated methods. Vancomycin Etest MICs were higher than those of other methods, whereas the MICs for daptomycin testing were comparable. Vancomycin MICs vary depending on the testing methodology.

Evaluation of the Etest method for susceptibility testing ofAspergillusspp. andFusariumspp. to three echinocandins

We performed Etest and broth microdilution (BMD) susceptibility testing of caspofungin, micafungin and anidulafungin against 67 clinical isolates of Aspergillus spp. and 10 Fusarium spp. Minimal effective concentrations (MECs) by BMD were read after 24 h of incubation at 35 degrees C and Etest MICs were read at 24 and 48 h. MECs < or =0.25 mg/l were obtained with caspofungin for all Aspergillus spp. tested but Etest MICs were < or =1 mg/l at 24 h. The agreement between caspofungin Etest and broth microdilution was good for all Aspergillus species tested (range 82.4-100%) except for A. niger and A. glaucus at 24 h of incubation. Micafungin and anidulafungin MEC and MIC results were lower than those of caspofungin (< or =0.015 mg/l) at 24 and 48 h for all Aspergillus tested. The agreement between the methods was excellent (100%) for micafungin and anidulafungin for all Aspergillus species tested. The three echinocandins were inactive against all isolates of Fusarium spp. showing MECs and MICs >8 mg/l. The Etest method could be a suitable procedure to test the susceptibility of most Aspergillus species to caspofungin, micafungin and anidulafungin; the best agreement was at 24 h.

In VivoEfficacy of Simulated Human Dosing Regimens of Prolonged-Infusion Doripenem against Carbapenemase- ProducingKlebsiella pneumoniae

Carbapenemase-producing Klebsiella pneumoniae (KPC) bacteria are rapidly becoming one of the most detrimental drug-resistant Gram-negative pathogens. Doripenem is the newest FDA-approved carbapenem that has the greatest in vitro potency against a wide range of Gram-negative organisms, including multidrug-resistant organisms. Previous work in an animal model has shown efficacy against Pseudomonas aeruginosa with MICs above the current breakpoints of susceptibility. The purpose of this study is to evaluate the efficacy of 1-g and 2-g dose prolonged infusions of doripenem against KPC isolates in both an immunocompetent and neutropenic murine thigh model. Seven clinical KPC isolates (broth microdilution [BMD] MIC range, 4 to 32 μg/ml; Etest MIC range, 3 to >32 μg/ml) were used. After infection, groups of mice were administered doripenem doses previously shown to simulate the exposures observed in humans after the administration of 1 or 2 g every 8 h as a 4-h infusion. In immunocompromised mice, 1- and 2-g doses of doripenem achieved bacteriostasis against isolates with MICs up to and including 8 μg/ml and 16 μg/ml, respectively. In immunocompetent animals, statistically significant reductions in the number of CFU were observed with overall decreases of approximately 1 log (P < 0.05). While carbapenemase-producing Klebsiella pneumoniae continues to decrease our meager supply of active agents, the ability of doripenem to produce CFU reductions in the presence of white blood cells (WBCs) using humanized exposures suggests the potential utility of this agent in combination against this increasingly problematic pathogen.

Comparison of Broth Microdilution, Agar Dilution, and Etest for Susceptibility Testing of Doripenem against Gram-Negative and Gram-Positive Pathogens

The susceptibility of 513 clinical isolates to doripenem was determined by broth microdilution, agar dilution, and Etest. Overall agreements for Etest and agar dilution MIC values compared to reference broth microdilution at +/-1 log(2) dilution were 88% and 94%, respectively. Etest MIC values demonstrated 98% agreement within +/-2 log(2) dilutions compared to the reference broth microdilution method.

Comparison of daptomycin Etest MICs on Mueller Hinton, IsoSensitest and brain heart infusion agars from Europe against 20 Staphylococcus aureus isolates

The Etest manufacturer (bioMerieux) recommends Mueller-Hinton agar (MHA) with calcium concentration of 25-40mg/L for daptomycin testing. A two phase study was performed to evaluate the effect of European agars on daptomycin Etest MICs. Broth microdilution (BMD) testing was performed and compared to Etest MICs for a challenge set of 20 Staphylococcus aureus with daptomycin MICs near the susceptible breakpoint and S. aureus 29213. In the first phase, Etest MICs were determined using agar plates prepared from MHA, IsoSensitest Agar (ISA) and Brain-Heart Infusion Agar (BHIA) and supplemented with various levels of calcium. In the second phase, Etest MICs were determined using commercially prepared MHA from Mast (MST), BD, Oxoid (OX), BioRad (BR), bioMerieux (BM) and E&O (EO) and ISA plates from OX, EO and MST. The calcium concentration of each agar was determined using ion selective electrode methodology. The best correlation of Etest and BMD MICs was obtained in phase 1 using MHA with 42.8mg/L calcium and in phase 2 with BD MHA. The phase 2 MHA calcium concentration ranged from 19.2 to 63.6mg/L. 87.5% of strains with BMD MICs of 1mg/L, had Etest MICs of 1.5 or 2mg/L using MHA with recommended calcium levels. Etest MICs using BHIA and ISA were higher than the BMD MICs and therefore are not recommended for use with the daptomycin Etest. The high percentage of major errors using Etest in this study, even with use of the most optimal medium, suggests that Etest MICs of 1.5 or 2mg/L using MHA should be retested by BMD.

Staphylococcus aureus with reduced glycopeptide susceptibility in Liverpool, UK

To investigate if colonization with heterogeneous glycopeptide-intermediate Staphylococcus aureus (hGISA) is associated with hGISA bacteraemia. Isolates of methicillin-resistant S. aureus (MRSA) from blood cultures and from swabs to detect MRSA colonization were screened for reduced susceptibility to glycopeptides by an agar incorporation method. Isolates detected by this screen were tested for glycopeptide resistance by MacroEtest, standard MIC Etest methods and population analysis profile-AUC (PAP-AUC) analysis. S. aureus isolates with and without reduced glycopeptide susceptibility were characterized by PFGE and spa typing. MRSA isolates with reduced susceptibility to glycopeptides, as identified by the MacroEtest method, were detected in the colonization screens of 86 of 2550 MRSA-positive patients. The isolates were confirmed by Etest MIC and PAP-AUC analysis as hGISA. A total of 82/86 of the hGISA colonizing isolates were EMRSA-16 by PFGE; the remainder were EMRSA-15. Bacteraemia with hGISA was identified in five patients during the study period; two isolates were EMRSA-16 and three were EMRSA-15. hGISA colonization could not be linked to hGISA bacteraemia and hGISA bacteraemia could not be linked to hGISA colonization. Four of the five hGISA bacteraemias developed following teicoplanin therapy for a central venous catheter-associated MRSA bacteraemia. Laboratory strategies to reduce morbidity associated with hGISA should focus on testing for hGISA in bacteraemic (rather than colonizing) MRSA isolates in patients with recurrent S. aureus bacteraemia following glycopeptide exposure.

Performance of Fusidic Acid (CEM-102) Susceptibility Testing Reagents: Broth Microdilution, Disk Diffusion, and Etest Methods as Applied to Staphylococcus aureus

Fusidic acid (CEM-102) is an established antistaphylococcal agent that has been used in clinical practice for more than 4 decades. The activity of fusidic acid against 778 isolates of Staphylococcus aureus collected from U.S. (53.8% were methicillin-resistant S. aureus [MRSA]) and Canadian (46.5% were MRSA) medical centers was assessed to determine the intermethod accuracy of the Clinical and Laboratory Standards Institute (CLSI) and Etest methods. Broth microdilution MIC results were compared by scattergram analysis to zone diameters around commercially available 5- and 10-microg disks. Acceptable correlation (r = 0.74 to 0.76) was observed for the two disk concentrations, and applying breakpoints of < or = 1 microg/ml (> or = 22 mm) for susceptibility (S) and > or = 4 microg/ml (< or = 19 mm) for resistance (R) provided 99.9% absolute intermethod categorical agreement. Reference CLSI MIC versus Etest MIC results (r = 0.77; 728 strains) showed 55.4% identical results and agreement of 99.7% +/- one log2 dilution. The diagnostic susceptibility testing reagents (including Etest) for fusidic acid (CEM-102) performed at an excellent level of intermethod agreement for the proposed breakpoint criteria.

Comparison of Anidulafungin MICs Determined by the Clinical and Laboratory Standards Institute Broth Microdilution Method (M27-A3 Document) and Etest for Candida Species Isolates

Anidulafungin Etest and CLSI MICs were compared for 143 Candida sp. isolates to assess essential (within 2 log(2) dilutions) and categorical agreements (according to three susceptibility breakpoints). Based on agreement percentages, our data indicated that Etest is not suitable to test anidulafungin against Candida parapsilosis and C. guilliermondii (54.4 to 82.4% essential and categorical agreements) but is more suitable for C. albicans, C. glabrata, C. krusei, and C. tropicalis (87.9 to 100% categorical agreement).

Caspofungin Etest Endpoint for Aspergillus Isolates Shows Poor Agreement with the Reference Minimum Effective Concentration

The Clinical and Laboratory Standards Institute (CLSI) M38-A2 reference broth microdilution (BMD) method for the antifungal susceptibility testing of filamentous fungi now includes guidelines for testing echinocandin activity using the minimum effective concentration (MEC) as the endpoint measurement. In this study, we compared the caspofungin Etest MIC on RPMI agar and Mueller-Hinton agar (supplemented with glucose and methylene blue [MGM]) to the BMD MEC for 345 clinical Aspergillus isolates, including A. flavus, A. fumigatus, A. nidulans, A. niger, and A. terreus. The essential agreement (+/-1 log(2) dilution) of the Etest on MGM and RPMI agar with the reference BMD MEC was 18 and 26%, respectively. The geometric mean values for BMD MEC and MGM Etest were 0.137 and 0.024 microg/ml, respectively, and the geometric mean values for BMD and RPMI agar were 0.128 and 0.031 microg/ml, respectively. Comparatively, 91% of paired MGM and RPMI Etest results were within 2 log(2) dilutions of each other and consistently produced clearly defined endpoints. In conclusion, the caspofungin Etest MIC, like the BMD MEC, is a reproducible endpoint but is markedly lower than the reference BMD. In anticipation of susceptibility breakpoint assignments, optimization studies will be required to improve the concordance of these two assays so that the potential for underreporting echinocandin resistance in Aspergillus is mitigated.

Evidence for daptomycin Etest lot-related MIC elevations for Staphylococcus aureus

MIC testing was performed simultaneously by Etest and broth microdilution (BMD) on 587 Staphylococcus aureus isolates submitted by local laboratories to a reference laboratory for confirmatory testing (May 2005 to July 2008). Testing bias was assessed for Etest to BMD MIC ratios. Categoric and essential agreement, very major (BMD nonsusceptible, Etest susceptible), and major (BMD susceptible, Etest nonsusceptible) errors were evaluated. Agar and broth calcium concentrations were consistent with current Clinical and Laboratory Standards Institute and manufacturer recommendations. There was a consistent bias for higher Etest MIC values compared with BMD. Ratios ranged from 0.25 to 4 (average 1.3), with substantial variability noted among the 8 different Etest lots tested. Overall, 6% of all ratios were >2.0. Categoric agreement and essential agreement among the 8 Etest lots ranged from 73% to 96% and 74% to 100%, respectively; very major errors ranged from 3% to 9%, and major errors ranged from 6% to 35%. However, most of the discrepancies were limited to 3 Etest lots.

Influence of testing methodology on the tigecycline activity profile against presumably tigecycline-non-susceptible Acinetobacter spp.

To compare the tigecycline activity profile against Acinetobacter spp. by Etest versus broth microdilution in isolates with high Etest MIC. Acinetobacter spp. isolates with tigecycline MICs of >or=0.5 mg/L determined by commercially developed Etests strips (January 2006 to July 2007) in five Spanish hospitals were considered. Values were rounded to the nearest upper double-dilution. Susceptibility by broth microdilution following CLSI (formerly NCCLS) recommendations, as the reference method, was determined in a central laboratory. BSAC breakpoints were used: susceptible <or=1 mg/L; intermediate = 2 mg/L; and resistant >2 mg/L. One hundred and forty-eight isolates were collected: 12 isolates with a tigecycline Etest MIC of 0.5 mg/L, 14 with 1 mg/L, 86 with 2 mg/L, 31 with 4 mg/L and 5 with 8 mg/L. Isolates with Etest MICs of 0.5-1 mg/L showed the same values by broth microdilution. Among isolates with Etest MICs of 2 mg/L, only 5.8% of strains showed the same value by both methods (88.4% showed values that were one or two dilutions lower by microdilution). None of the 36 isolates with Etest MICs of 4-8 mg/L showed the same value by both methods, with values at least two dilutions lower by microdilution. Weak correlation (R = 0.238; P <or= 0.001) was found between both methods. All 26 Etest susceptible isolates, 80/86 (93.0%) Etest intermediate and 32/36 (88.9%) Etest resistant strains were susceptible by microdilution. Caution should be taken in interpreting Etest MICs of >or=2 mg/L for Acinetobacter spp. since strains with Etest MICs of 2-4 mg/L are susceptible when tested by microdilution. False non-susceptibility by Etest may exclude tigecycline as a therapeutic option in a field where multiresistance is the rule.

Vancomycin MICs for Staphylococcus aureus Vary by Detection Method and Have Subtly Increased in a Pediatric Population Since 2005

Vancomycin MICs for Staphylococcus aureus isolates in a pediatric hospital with a high rate of staphylococcal infections were examined for any increase over a 7-year period. A broth microdilution scheme allowed direct comparison of the MICs generated by this method to MICs generated by Etest. MICs generated by both methods were determined with the same inoculum suspension. One hundred sixty-five S. aureus isolates were selected on the basis of the patients having been bacteremic or having received vancomycin as the definitive therapy for their infections. Of the 165 isolates, 117 were methicillin-resistant S. aureus and 48 were methicillin-susceptible S. aureus. Forty-seven were acquired in the hospital (nosocomial), 56 were community acquired, and 62 were community onset-health care associated. All but one isolate tested by broth microdilution had MICs of < 1.0 microg/ml, while 96% of these same isolates tested by Etest had MICs of > or = 1 microg/ml. A significant increase in MICs that occurred after study year 4 (2004 to 2005) was demonstrated by the Etest (P < 0.00007) but not by broth microdilution. MICs were not different for isolates of community or health care origin, regardless of methodology. The proportion of isolates with Etest MICs of < 1 and > or = 1 microg/ml between children with bacteremia for < or = 5 days and > 5 days (P = 0.3) was not different. We conclude that MICs for pediatric isolates have increased slightly since 2005 and therapeutic decisions based on vancomycin MICs need to be made by considering the methodology used.

Evaluation of the Etest GRD for the detection of Staphylococcus aureus with reduced susceptibility to glycopeptides

Continued glycopeptide-selective pressure has led to non-susceptible strains of Staphylococcus aureus including heterogeneously vancomycin-intermediate S. aureus (hVISA). The gold standard for identification of hVISA is the population analysis profile area under the curve ratio (PAP-AUC), though this method is time-consuming and labour-intensive. The objective of this study was to compare a new method for detection of hVISA, the Etest GRD, to PAP-AUC and to macro Etest. One hundred clinical hVISA and 50 clinical fully vancomycin-susceptible S. aureus (VSSA), confirmed by PAP-AUC, were evaluated. Microtitre and Etest MICs were determined by standard testing procedures on all isolates. Macro Etest was performed according to referenced procedures. The Etest GRD was tested using a 0.5 McFarland standard on Mueller-Hinton agar + 5% blood and read at 24 and 48 h. If either the vancomycin or the teicoplanin end of the GRD strip was >or=8 and the vancomycin Etest was <or=4, the isolate was considered hVISA. Vancomycin MIC(50)/MIC(90) for hVISA and VSSA was 1.5/2 mg/L and 1/1.5 mg/L, respectively, by Etest and vancomycin MIC(50)/MIC(90) for hVISA and VSSA was 1/2 mg/L for both by microtitre; MIC values for hVISA being significantly higher (P <or= 0.023). At 24 h, the Etest GRD was 77% sensitive and 98% specific, and at 48 h, it was 93% sensitive and 82% specific compared with PAP-AUC. Macro Etest was 83% sensitive and 94% specific at 48 h. Etest GRD was simple to perform and may be feasible for clinical microbiology laboratories. This test may be useful for clinical detection of hVISA.

Comparison of the Etest and the routine multi-disc agar diffusion susceptibility of &lt;i&gt;Staphylococcus&lt;/i&gt; species

Aims: The present study, tend to evaluate the validity and accuracy of Etest as a method for performing in-vitro antimicrobial susceptibility testing of Staphylococcus with comparison to the routine multi disc agar diffusion. This is because the Etest susceptibility method is not yet known as a rapid, simple reliable technique in developing countries as it combine the functions of both dilution and diffusion technique. Materials and methods: Ninety-seven Staphylococcus aureus and eightythree Staphylococcus epidermidis isolates were obtained from wound samples and identified according to standard morphological and biochemical methods. The antibiotics susceptibility patterns were determined both by agar disc diffusion and Etest methods in accordance to NCCLS (1997) criteria and manufacturer (AB Biodisk Sweden) respectively. Results: On the Etest strips, Staph aureus was 83.5% sensitive to ciprofloxacin, 52.6% to gentamicin, 48.5% to ampicillin and 8.2% to chloramphenicol while on the multi-disc agar diffusion plates 80.4% of Staph aureus were sensitive to ciprofloxacin, 49.5% to gentamicin, 39.2% to ampicillin and 12.4% to chloramphenicol.. On the Etest strips, 80.7% of Staph epidermidis were sensitive to ciprofloxacin, 34.9% to gentamicin, 25.3% to ampicillin and 15.7% to chloramphenicol while on the multi- disc agar diffusion plates 89.2% of Staph epidermidis were sensitive to ciprofloxacin, 34.9% to gentamicin, 25.3% to ampicillin and 32.5% to chloramphenicol. Conclusion: The sensitivity patterns between the two methods were essentially similar, however, the Etest method clearly demonstrated intermediate sensitivities which to an extent were absent in routine multi-disc agar diffusion method. Most of the isolates Etest MICs clustered around the sensitive and resistance break points. Etest also demonstrated the MIC and diffusion results on the same strips. Keywords: antibiotic resistance, antimicrobial, gram-positive, chemotherapy.Sudan Journal of Medical Sciences Vol. 3 (2) 2008: pp.121-126

In Vitro Activity of Tigecycline against Gram-Positive and Gram-Negative Pathogens as Evaluated by Broth Microdilution and Etest

The current surveillance establishes the activity profile of tigecycline against recent clinical U.S. isolates of target pathogens. Findings from a distributed surveillance that utilized Etest yielded a tigecycline activity profile that varied from that observed in a separate centralized broth microdilution (BMD) surveillance (D. C. Draghi et al., Poster D-0701, 46th Intersci. Conf. Antimicrob. Agents Chemother., San Francisco, CA). Differences were noted among Acinetobacter spp. and Serratia marcescens and, to a lesser extent, with Streptococcus pyogenes. To address whether these differences were due to discordance in testing methodology or to variations among the analyzed populations, isolates from the current surveillance were concurrently tested by BMD and Etest. In all, 1,800 Staphylococcus aureus, 259 S. pyogenes, 226 Streptococcus pneumoniae, 93 Enterococcus faecalis, 1,356 Enterobacteriaceae, and 227 Acinetobacter baumannii strains were evaluated. Tigecycline had potent activity by BMD, with >99.6% susceptibility (%S) observed for all pathogens with interpretive criteria, excluding Enterobacter cloacae (98.3% S) and E. faecalis (86.0% S), and MIC(90)s ranged from 0.03 mug/ml (S. pyogenes/S. pneumoniae) to 1 mug/ml (Enterobacteriaceae/A. baumannii). Similar profiles were observed by Etest, with the exception of A. baumannii, although for most evaluated pathogens Etest MICs trended one doubling-dilution higher than BMD MICs. Major or very major errors were infrequent, and a high degree of essential agreement was observed, excluding A. baumannii, S. marcescens, and S. pneumoniae, for which >/=4-fold differences in MICs were observed for 29, 27.1, and 34% of the isolates, respectively. Further analysis regarding the suitability of the tigecycline Etest for testing S. marcescens, Acinetobacter spp., and S. pneumoniae is warranted.

Molecular Epidemiology of Methicillin-Resistant Staphylococcus aureus Bloodstream Isolates in Urban Detroit

To gain a better understanding of epidemiology of resistance in Staphylococcus aureus, we describe the molecular epidemiology of methicillin-resistant Staphylococcus aureus bloodstream isolates in urban Detroit. Bloodstream isolates from July 2005 to February 2007 were characterized. Two hundred ten bloodstream isolates from 201 patients were evaluated. Patient characteristics were as follows: median age, 54 years; 56% male; and 71% African-American. Seventy-six percent of infections were health care associated, with 55% being community-onset infections and 21% hospital acquired, and 24% were community associated. The most common sources were skin/wound (25%), central venous catheters (24%), unknown source (20%), and endocarditis (9%). Ninety percent and 5% of isolates had a MIC of vancomycin of <or=1.0 mg/liter, using automated dilution testing and E-test, respectively. Six percent of isolates showed heteroresistance to vancomycin, all occurring with isolates having a vancomycin E-test MIC of >or=1.5 mg/liter. Results of pulsed-field gel electrophoresis showed 17 strain types. The predominant strains were USA100 (104 isolates) and USA300 (74 isolates). Forty-nine percent of the isolates had staphylococcal cassette chromosome mec II, and 56% had agr II. All USA300 isolates were positive for the Panton-Valentine leukocidin toxin genes and agr I. Forty-seven percent of USA300 bloodstream infections were health care associated (35% community onset and 12% hospital onset). USA300 strains were more common in injection drug users with skin/wound as the predominant source of infection. Thirty percent of the USA100 strains were closely related to vancomycin-resistant Staphylococcus aureus isolates. The results of this study show that vancomycin MICs using automated dilution testing with Vitek-2 and E-test were highly discordant. Most methicillin-resistant S. aureus strains causing bacteremia are health care associated, commonly have MICs of vancomycin that are high within the susceptible range are not detected by routine automated dilution testing, and have significant diversity of molecular characteristics. USA100 strains that are closely related to vancomycin-resistant S. aureus (VRSA) isolates and USA300 strains are common as causes of both hospital and community-onset infection. Infection control measures should focus not only on prevention of the spread of community strains in the hospital but also prevention of the spread of hospital strains associated with VRSA into the community.