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PubMed2001

Performance of the chromogenic medium CHROMagar Staph Aureus and the Staphychrom coagulase test in the detection and identification of Staphylococcus aureus in clinical specimens.

CHROMagar Staph aureus (CSAM) (CHROMagar Microbiology, Paris, France) is a new chromogenic medium designed to enable detection of colonies of Staphylococcus aureus by their pink color. A total of 775 specimens were cultured in parallel on CHROMagar Staph aureus and conventional media. Among the 267 S. aureus strains recovered on at least one medium, 263 were isolated on CSAM medium (sensitivity, 98.5%), and 245 (sensitivity, 91.8%) were isolated on conventional media. The specificity of presumptive identification of S. aureus on the basis of pink colony color on CSAM medium was 97% (493 of 508). This specificity increased to 100% when coagulase detection with the Staphychrom coagulase test was added and to 98.8% when S. aureus surface components were detected by agglutination in the Pastorex Staph Plus test. Susceptibility testing of 67 S. aureus strains, performed in parallel on pink CSAM colonies and on colonies grown on blood agar, gave similar results. Thus, rapid and accurate recognition and identification of S. aureus isolates were achieved with CSAM as the primary isolation medium, followed by the staphylocoagulase Staphychrom test. Antimicrobial susceptibility testing (disk-diffusion method or ATB STAPH System) can be performed directly on pink CSAM colonies.

Chromogenic Compounds↗
PubMed2006

[Community-acquired methicillin-resistant Staphylococcus aureus disseminated disease].

A 21 year old man, previously healthy, presented with subcutaneous nodes consistent with gummas. Ultrasonography disclosed multiple subcutaneous abscesses and images suitable with piomiositis, pleural and pericardium effusion. A puncture-aspirate with fine-needle was performed and produced purulent material, with isolate of Staphylococcus aureus. Antimicrobial susceptibility testing by disk diffusion showed resistant to cefalotin, erythromycin and clindamycin, and susceptibility to trimethoprim-sulfamethoxazole, ciprofloxacin and rifampicin. Methicilin-resistance was confirmed by Staphyslide agglutination testing (Biomérieux). The patient was treated with ciprofloxacin and rifampicin during four weeks, with a good clinical response. The frequency of CA-MRSA infections is increasing, and these are reported in patients without identified predisposing risks leading to failure on empiric therapy for community infections presumed to be due to staphylococcal agents.

Abscess↗
PubMed1995

Evaluation of E-Test for determination of antimicrobial MICs for Pseudomonas aeruginosa isolates from cystic fibrosis patients.

We determined the E-Test and National Committee for Clinical Laboratory Standards standardized agar dilution MICs of ceftazidime, ciprofloxacin, piperacillin, and tobramycin for Pseudomonas aeruginosa during tests of 100 rough and mucoid P. aeruginosa isolates from cystic fibrosis patients. The levels of agreement (+/- 1 log2 dilution) between quantitative E-Test and agar dilution MIC results were 80, 97, 73, and 89% for ceftazidime, ciprofloxacin, piperacillin, and tobramycin, respectively. Comparison of the results after converting the MIC data to qualitative categories (susceptible, intermediate, and resistant) yielded levels of agreement of 84, 96, 88, and 93% for the same agents, respectively. Of the 39 qualitative discrepancies, 36 were minor and 3 were very major. We conclude that use of the E-Test is easier and more practical than use of the agar dilution method for most laboratories and that the E-Test furnishes results which are at least as accurate as those obtained by the agar dilution method. However, the higher cost of the E-Test method would likely discourage most laboratories from selecting it over disk diffusion for routine antimicrobial susceptibility testing of P. aeruginosa isolates from cystic fibrosis patients.

Cystic Fibrosis↗
PubMed2004

Antimicrobial susceptibility testing of Acinetobacter spp. by NCCLS broth microdilution and disk diffusion methods.

Although both broth microdilution (BMD) and disk diffusion (DD) are listed by NCCLS as acceptable methods for testing Acinetobacter spp. for antimicrobial susceptibility, few studies have compared the results generated by the two methods. We tested 196 isolates of Acinetobacter spp. from nine U.S. hospitals and from the Centers for Disease Control culture collection by using BMD and DD and clinically appropriate antimicrobial agents. Categorical results for amikacin, ciprofloxacin, gatifloxacin, gentamicin, imipenem, levofloxacin, meropenem, tobramycin, and trimethoprim-sulfamethoxazole were comparable for the two methods: there was only one very major (VM) error, with tobramycin, and only one major (M) error, with meropenem, when DD results were compared with BMD results. However, VM errors were frequent with the beta-lactams and beta-lactam-beta-lactam inhibitor combinations, while M errors were often observed with tetracyclines. For BMD, tests frequently exhibited subtle growth patterns that were difficult to interpret, especially for beta-lactams. If subtle growth (i.e., granular, small button, or "starry" growth) was considered positive, error rates between BMD and DD were unacceptably high for ampicillin-sulbactam (VM error, 9.8%; minor [m] error, 16.1%), piperacillin (VM error, 5.7%; m error, 13.5%), piperacillin-tazobactam (VM error, 9.3%; m error, 12.9%), ceftazidime (VM error, 6.2%; m error, 11.4%), cefepime (VM error, 6.2%; m error, 13.0%), cefotaxime (m error, 21.2%), ceftriaxone (m error, 23.3%), tetracycline (M error, 11.4%; m error, 32.1%), and doxycycline (M error, 2.6%). When subtle growth patterns were ignored, the agreement still did not achieve acceptable levels. To determine if the problems with BMD testing occurred in other laboratories, we sent frozen BMD panels containing beta-lactam drugs and nine isolates to six labs with experience in performing BMD and DD. Among these laboratories, cefepime MICs ranged from < or =8 to > or =32 microg/ml for four of the nine strains, confirming the problem in interpreting BMD results. Discrepancies between the categorical interpretations of BMD and DD tests were noted primarily with cefepime and piperacillin, for which the BMD results were typically more resistant. Clinical laboratories should be aware of these discrepancies. At present, there are no data to indicate which method provides more clinically relevant information.

Acinetobacter↗
PubMed1991

Comparison of the E Test to agar dilution, broth microdilution, and agar diffusion susceptibility testing techniques by using a special challenge set of bacteria.

The E Test (AB Biodisk, Solna, Sweden) is a new method for performing antimicrobial susceptibility tests. It consists of an impervious carrier (5- by 50-mm strip) with a predefined antimicrobic gradient which is placed on an inoculated agar plate and processed like a disk diffusion test. Results are generated directly as MICs from a continuous concentration gradient covering 15 twofold dilutions, and MICs are read where the edge of the inhibition zone intersects the strip. We compared the E Test with disk diffusion, broth microdilution, and agar dilution tests by using a challenge set of 195 gram-positive and gram-negative bacteria for 14 antimicrobial agents. Also, disk diffusion, broth microdilution, and agar dilution tests were compared with each other. All test method comparisons gave greater than 94% agreement for the category of susceptibility. The E Test category agreement with disk diffusion and broth microdilution was 95.1%, and with agar dilution it was 95.2%. The E Test results were as reliable as the results obtained by the standard antimicrobial susceptibility testing methods.

Agar↗
PubMed2001

Study on antimicrobial susceptibility of bacteria causing neonatal infections: a 12 year study (1987-1998).

OBJECTIVE: The method of Manual of Clinical Microbiology was used to identify bacteria. We investigated the epidemiological characteristics of bacterial agents and their antimicrobial susceptibility as empirical treatment for neonatal infections. Disk diffusion tests were also done for antimicrobial susceptibility. RESULTS: From January 1987 to December 1998, 2,244 bacterial strains were isolated in our neonatal ward. The first three predominant species were Staphylococcus epidermidis (23.9%), Staphylococcus saprophyticus (19.9%) and Escherichia coli (12.6%) in group I (infections acquired outside of hospital). Escherichia coli, Klebsiella and Pseudomonas aeruginosa accounted for 18%, 15.2% and 12.6% respectively in group II (nosocomial infections). The sensitivity rates of those antimicrobials that are seldom used for newborns were found to be higher, while the resistant rates of the commonly used antimicrobial drugs have increased significantly. The resistant rates of bacterial isolate from group II to antimicrobial agents including penicillin and ampicillin were significantly higher than those isolated from group I (p<0.05)The sensitivity rate was 82.2% (717/833) by using amikacin only, when combined with penicillin, rose to 89%(741/833). CONCLUSIONS: Gram-negative bacteria were mainly responsible for nosocomial infections of neonates in our hospital. Infections acquired outside the hospital were mainly caused by Gram-positive bacteria. Nosocomial pathogens produced drug resistance easily. Combination of amikacin and penicillin can be recommended as the initial antibiotics for treatment of neonatal infections.

Anti-Bacterial Agents↗
PubMed1977

Disk diffusion susceptibility testing of Nocardia species.

The effectiveness of 13 antimicrobial agents against 51 clinical isolates of Nocardia was determined with use of agar dilutions and disk diffusion method. Amikacin inhibited less than 90% of isolates and, like the other aminoglycosides, showed good correlation between minimal inhibitory concentrations and sizes of zones of inhibition around the disks. Both sulfisoxazole and trimethoprim-sulfamethoxazole were very active, although they required a 2- to 3-log lower inoculum for demonstration of susceptibility. Results with the two sulfa disks were variable, but they did allow distinction between sensitive and intermediate strains. All of the isolates of Nocardia were inhibited by 6.3 microng of minocycline; however, the degree of susceptibility could not be determined by zone diameters. Only two-thirds of these clinical isolates of Nocardia grew rapidly enough to be assayed by either susceptibility method.

Anti-Bacterial Agents↗
PubMedDate not supplied

Antibiotic susceptibility of Stenotrophomonas (Xanthomonas) maltophilia: comparative (NCCLS criteria) evaluation of antimicrobial drugs with the agar dilution and the agar disk diffusion (Bauer-Kirby) tests.

Ninety-six clinical isolates of Stenotrophomonas maltophilia were examined with the agar dilution method for susceptibility to 19 antimicrobial drugs. Doxycycline, cotrimoxazole, timentin, ofloxacin, fosfomycin, and piperacillin + tazobactam, in that order, inhibited the majority of strains. All isolates were resistant to nitrofurantoin. Concurrent disk susceptibility (Bauer-Kirby method) testing, using currently valid NCCLS interpretative criteria for Pseudomonas aeruginosa, uncovered a significant incidence of very major (category I), major (category II), and minor (categories III and IV) discrepancies for aminoglycosides, cephalosporins, chloramphenicol, and piperacillin + tazobactam and ticarcillin + clavulanic acid. Therefore, new interpretative criteria indicative of intermediate (I) susceptibility of S. maltophilia to these various antibiotics were proposed. In addition, new intermediate susceptibility criteria were proposed for the two beta-lactam-beta-lactamase inhibitor combinations. It was recommended to exclude ciprofloxacin from test batteries against this microorganism due to the wide scatter of minimal inhibitory concentration values and diameters of inhibition zones; the same was true for polymyxin B. It is hoped that the proposed modified, species-specific criteria will improve the clinical utility of laboratory-generated disk antibiograms with respect to the inherently multiple antibiotic-resistant, opportunistic pathogen S. maltophilia.

Anti-Bacterial Agents↗
PubMed1983

Susceptibility of Mycobacterium fortuitum to antibacterial agents as evaluated by disk diffusion and agar dilution methods.

The susceptibility of 34 strains of M. fortuitum to several antimicrobial agents was comparatively tested by the disk diffusion and the agar dilution methods. The results obtained by these two methods essentially showed that the microorganism was fully sensitive to amikacin and kanamycin and only partially sensitive to sisomicin and rifampicin. Streptomycin, erythromycin and cephotaxime were inactive. The disk diffusion method seems advisable as a rapid screening procedure for antibiotic susceptibility testing M. fortuitum.

Antibiotics, Antitubercular↗
PubMed2005

Intra- and interlaboratory performance of antibiotic disk-diffusion-susceptibility testing of bacterial control strains of relevance for monitoring aquaculture environments.

In the course of an international research project on hazard analysis of antimicrobial resistance in SE Asian aquaculture environments, 2 European Union and 3 SE Asian laboratories attempted to harmonize a procedure for antimicrobial agent susceptibility testing based on disk diffusion (DD). For this purpose, a selected panel of 10 bacterial control strains of relevance for monitoring warm-water aquaculture environments was sent by the central laboratory to the other participating laboratories. In each laboratory, 10 independently replicated DD determinations of each control strain to 6 antibiotics were performed using Iso-Sensitest Agar (ISA) according to a standard operating procedure (SOP); in total, this study thus yielded 300 data sets for all 5 laboratories. At the end of the study, strain authenticity of subcultures of the control strains used by the respective participating laboratories was verified by the central laboratory. Based on the arithmetic mean of 10 inhibition-zone diameter measurements and standard deviation (SD), intralaboratory SD variations ranged from 0 to 2 mm when 79% of the recorded data sets were considered. In 8% of the data sets, the SD value exceeded 4 mm, which in most cases could be attributed to the fact that the data points for a given strain-disk combination were not normally distributed in one of the laboratories. At the interlaboratory level, 81% of the SD values based on global averaging of 50 data points per strain-disk combination were situated in the 0 to 5 mm range. Comparison with a minimal data set from literature of DD testing performed with Mueller-Hinton (MH) medium indicated that the use of either ISA or MH medium in DD testing has a limited impact on the method's precision among different laboratories. In conclusion, the current study has provided a validated SOP to promote the coordination and harmonization of DD-susceptibility methodologies for aquaculture-associated organisms at an international level. As one of the main action items for the future, new interpretive breakpoints should be specifically designed and validated for aquaculture drugs and organisms.

Anti-Bacterial Agents↗
PubMed2005

Patterns of antimicrobial resistance observed in Escherichia coli isolates obtained from domestic- and wild-animal fecal samples, human septage, and surface water.

A repeated cross-sectional study was conducted to determine the patterns of antimicrobial resistance in 1,286 Escherichia coli strains isolated from human septage, wildlife, domestic animals, farm environments, and surface water in the Red Cedar watershed in Michigan. Isolation and identification of E. coli were done by using enrichment media, selective media, and biochemical tests. Antimicrobial susceptibility testing by the disk diffusion method was conducted for neomycin, gentamicin, streptomycin, chloramphenicol, ofloxacin, trimethoprim-sulfamethoxazole, tetracycline, ampicillin, nalidixic acid, nitrofurantoin, cephalothin, and sulfisoxazole. Resistance to at least one antimicrobial agent was demonstrated in isolates from livestock, companion animals, human septage, wildlife, and surface water. In general, E. coli isolates from domestic species showed resistance to the largest number of antimicrobial agents compared to isolates from human septage, wildlife, and surface water. The agents to which resistance was demonstrated most frequently were tetracycline, cephalothin, sulfisoxazole, and streptomycin. There were similarities in the patterns of resistance in fecal samples and farm environment samples by animal, and the levels of cephalothin-resistant isolates were higher in farm environment samples than in fecal samples. Multidrug resistance was seen in a variety of sources, and the highest levels of multidrug-resistant E. coli were observed for swine fecal samples. The fact that water sample isolates were resistant only to cephalothin may suggest that the resistance patterns for farm environment samples may be more representative of the risk of contamination of surface waters with antimicrobial agent-resistant bacteria.

Animals↗
PubMed2004

Molecular epidemiology and control of nosocomial methicillin-resistant Staphylococcus aureus infection in a teaching hospital.

BACKGROUND AND PURPOSE: Nosocomial methicillin-resistant Staphylococcus aureus (MRSA) infection is difficult to control. Due to a dramatic increase in the nosocomial MRSA infection rate at our hospital from 2000 to 2001, this study was conducted to identify the source of these infections and the effectiveness of control measures. METHODS: 179 healthcare workers (HCWs) were screened for carriage of MRSA. Starting in April 2001, all patients with MRSA infection or colonization were put in strict contact and cohort isolation. The bacterial isolates of HCW carriers and patients with MRSA infection from April 2001 to September 2001 were subjected to antimicrobial susceptibility testing by disk-diffusion method and molecular typing by pulsed-field gel electrophoresis (PFGE). RESULTS: Fifteen HCWs were found to be carriers of MRSA. They were all given topical mupirocin treatment. After these interventions, the nosocomial MRSA infection rate decreased from 1.23 to 0.53 per 1000 patient-days. All 61 MRSA isolates available for antimicrobial susceptibility testing and molecular typing were multidrug resistant. PFGE study revealed 2 predominant types, type C and type Y, comprising 36 and 12 isolates, respectively. CONCLUSIONS: The current study demonstrates the importance of measures to control nosocomial MRSA infections in hospitals that already have a high incidence of endemic MRSA infection. Elimination of carriage by healthcare workers, and strict contact and cohort isolation are the main effective measures.

Anti-Bacterial Agents↗
PubMed1987

Institution-specific versus published in vitro antimicrobial susceptibility data in making formulary decisions.

The results of susceptibility testing of 549 isolates of gram-negative organisms to 17 antimicrobial agents were compared with published reports of the sensitivity of those organisms to those agents. All gram-negative bacilli isolated from cultures obtained from hospitalized patients during a three-month period were preserved for antimicrobial sensitivity testing. Standard Kirby-Bauer disk diffusion susceptibility tests were performed using 17 broad-spectrum antimicrobial agents that either were included in the hospital formulary or were being considered for inclusion. Organisms were recorded as being sensitive or resistant to each drug, and the results were compared with the published results of in vitro sensitivity studies. When the results of actual antimicrobial sensitivity testing varied from published results, the discordant results were assigned a ranking of 1 to 4 based on the percentage difference. In 34 of 77 drug-organism pairs tested, the results of susceptibility testing differed by more than 10% cumulative susceptibility from published values; 26 of these represented instances in which the results of actual testing were at least 10% less than published values. For seven of the antimicrobial agents that were being considered for inclusion in the hospital formulary, results indicating unexpectedly suboptimal activity against institutional pathogens were a determinant in eliminating the agents from further consideration. In vitro testing of antimicrobial susceptibility of local pathogens can be a better method of predicting the susceptibility of such pathogens to new antimicrobial agents than relying on published susceptibility data. Pharmacy and therapeutics committees should consider testing prevalent institutional pathogens for susceptibility to all antimicrobial agents that are proposed additions to the formulary.

Aminoglycosides↗
PubMed1999

Combinations of vancomycin and beta-lactams are synergistic against staphylococci with reduced susceptibilities to vancomycin.

Evidence of synergism between combinations of vancomycin and beta-lactam antibiotics against 59 isolates of methicillin-resistant staphylococci (Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus haemolyticus) for which vancomycin MICs ranged from 1 to 16 microg/ml were tested by broth microdilution checkerboard, disk diffusion, agar dilution, and time-kill antimicrobial susceptibility tests. The combination of vancomycin and oxacillin demonstrated synergy by all test methods against 30 of 59 isolates; no antagonism was seen. Synergy with vancomycin was also found by modified disk diffusion testing for ceftriaxone, ceftazidime, cefpodoxime, and amoxicillin-clavulanate but not for aztreonam. Evidence of synergy correlated directly with vancomycin MICs. The efficacy of vancomycin given alone and in combination with nafcillin was tested in the rabbit model of experimental endocarditis caused by three clinical isolates of glycopeptide-intermediate-susceptible S. aureus (GISA) (isolates HIP5827, HIP5836, and MU50). Two of the GISA isolates (isolates MU50 and HIP5836) were extremely virulent in this model, with 27 of 42 (64%) animals dying during the 3-day trial. Therapy with either vancomycin or nafcillin given as a single agent was ineffective for animals infected with HIP5827 or MU50. However, the combination of vancomycin and nafcillin resulted in a mean reduction of 4.52 log10 CFU/g of aortic valvular vegetations per g compared to the reduction for controls for animals infected with HIP5827 and a reduction of 4. 15 log10 CFU/g for animals infected with MU50. Renal abscesses caused by HIP5827 were sterilized significantly better with the combination of vancomycin and nafcillin than by either treatment alone. We conclude that the combination of vancomycin and beta-lactams with antistaphylococcal activity is an effective regimen for the treatment of infections with clinical strains of staphylococci which demonstrate reduced susceptibility to glycopeptides.

Animals↗
PubMed1990

[Prevention and control of nosocomial infection caused by methicillin resistant Staphylococcus aureus in premature infant ward--prevention effect of "povidone iodine solution" wipe of neonatal skin].

In January 1983, a number of premature infants under management in the premature infants' unit of our hospital were found to have bacteremia due to Staphylococcus aureus. By the end of February of the same year, 4 of these infants, who had been treated in the same unit, developed impetigo. The S. aureus responsible for this condition was classified as type IV by a coagulase typing. In a subsequent antimicrobial susceptibility test using the disk diffusion method, this microorganism was found to be resistant to methicillin, erythromycin and lincomycin, and to be susceptible to tetracycline, chloramphenicol and cefmetazole, indicating that it was a methicillin resistant S. aureus (MRSA). Because the result from the coagulase typing agreed with the antimicrobial susceptibility pattern in all cases, we concluded that these cases represented nosocomial infection with MRSA. The source and route of the infection were investigated, and measures taken to prevent bacterial spread from carriers and to keep instruments and environments clean. Although the source of infection was not identified. Then, we tried wiping the body surface of the premature infants with an Isodine solution (10% PVP-I, 1:100 dilution) in order to prevent colonization of the microorganism on the body surface. With this application+, MRSA was no longer detected from the body surface of the premature infants, and no additional MRSA infection occurred in the premature infants' unit. Data collected for premature infants' managed at our hospital in the subsequent 6 years allows us to conclude that MRSA infection can be almost completely controlled by frequent surveys of carriers and appropriate body surface wiping with Isodine solution.(ABSTRACT TRUNCATED AT 250 WORDS)

Cross Infection↗
PubMed2004

Etiology, antimicrobial susceptibility profiles, and mortality associated with bacterial meningitis among children in Egypt.

PURPOSE: Surveillance for patients with meningitis is a high priority in order to determine the etiology of disease and design prevention strategies. This study presents data on the causes of bacterial meningitis among children <6 years of age treated in a network of hospitals throughout Egypt. METHODS: Training was provided to standardize the collection of clinical information and optimize recovery of bacterial pathogens. Bacterial isolates were tested for antimicrobial resistance patterns using Kirby Bauer disk diffusion, E-test and/or Beta-lactamase (BL) testing methods. RESULTS: Patients with culture-confirmed bacterial meningitis (228 children<6 years) were identified including 89 (39%) patients with H. influenzae (HI), 68 (30%) with Streptococcus pneumoniae (SP), 30 (13%) with N. meningitidis (NM), 18 (8%) with Mycobacterium tuberculosis (MTB) and 23 (10%) with other bacteria. The overall case fatality ratio was high (24%) and increased among children with TB meningitis (56%). The susceptibility for HI to ampicillin (AMP), chloramphenicol (C) and ceftriaxone (CRO) was 21%, 13%, and 100% respectively. The susceptibility for SP to C and CRO was 79% and 100%, respectively. CONCLUSIONS: HI and SP are the leading causes of bacterial meningitis among children in Egypt. The majority of HI strains tested were resistant to AMP or C suggesting the need for routine use of CRO as first line therapy. Among older children TB emerges as a significant cause of bacterial meningitis in Egypt.

Age Factors↗
PubMed1978

Antimicrobial susceptibility patterns of Streptococcus pneumoniae.

Fifty clinical isolates of Streptococcus pneumoniae received by the Streptococcus Laboratory of the Center for Disease Control from August 1976 through March 1977 and 50 pneumococcal strains retrieved from 13- to 16-year storage (originally isolated October 1961 through December 1964) were tested for susceptibility to 10 antimicrobial agents by disk-agar diffusion and agar dilution tests. No appreciable differences in susceptibility patterns were apparent between the two groups, and, except for one isolate, all were highly susceptible to every drug tested except gentamicin. This single isolate required higher drug concentrations to inhibit macroscopic growth and had corresponding decrements in zones of disk inhibition with penicillin, ampicillin, and cephalothin. An additional 43 pneumococci recently received from various areas of the United States and Canada were screened by a disk agar diffusion method for penicillin resistance. Four of these isolates had penicillin zone diameters <30 mm, and subsequent agar dilution test results showed that the penicillin minimum inhibitory concentrations were elevated with these organisms. Antimicrobial susceptibility patterns of pneumococci to antimicrobials other than penicillin and its analogs have not changed substantially in over a decade. However, due to the emergence of strains with decreased susceptibility to penicillin, the screening test for penicillin resistance in pneumococci, especially in isolates from spinal fluid and blood, could be clinically useful as an aid in selecting optimal therapy.

Cephalosporinase↗