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Biomedical subjects

P R Murray

Publications and source records attributed to P R Murray.

At least 19 recordsLinked to original sources

Use of molecular and reference susceptibility testing methods in a multicenter evaluation of MicroScan dried overnight gram-positive MIC panels for detection of vancomycin and high-level aminoglycoside resistances in enterococci.

Modified MicroScan gram-positive MIC no. 8 panels (PM-8) were analyzed for their improved ability to detect vancomycin resistance (VR) and high-level aminoglycoside resistance (HLAR) in enterococci. A validation study design that utilized selected challenge strains, recent clinical isolates, and reproducibility experiments in a multicenter format was selected. Three independent medical centers compared the commercial panels to reference broth microdilution panels (RBM) and Synergy Quad Agar (QA). Resistance was verified by demonstration of VR and HLAR genes by PCR tests. The study was conducted in three phases. (i) In the challenge phase (CP), two well-characterized sets of enterococci were obtained from the Centers for Disease Control and Prevention; one set contained 50 isolates for VR testing and one contained 48 isolates for HLAR testing. In addition, a set of 47 well-characterized isolates representing diverse geographic areas, obtained from earlier national surveillance studies, was tested at the University of Iowa College of Medicine (UICM). (ii) In the efficacy phase (EP), each laboratory tested 50 recent, unique clinical isolates by all methods. (iii) In the reproducibility Phase (RP), each laboratory tested the same 10 strains by all methods in triplicate on three separate days. All isolates from the EP were sent to the UICM for molecular characterization of vanA, -B, -C1, -C2-3, and HLAR genes. In the CP, the ranking of test methods by error rates (in parentheses; very major and major errors combined, versus PCR results) were as follows: for high-level streptomycin resistance (HLSR), QA (12.0%) > PM-8 (5.2%) > RBM (1.6%); for high-level gentamicin resistance (HLGR), RBM (3.7%) > PM-8 (3.1%) > QA (2.6%); and for VR, RBM = QA (3.0%) > PM-8 (1.2%). In the EP, agreement between all methods and the reference PCR result was 98.0% for HLSR, 99.3% for HLGR, and 98. 6% for VR. In the RP, the percentages of results +/- 1 log2 dilution of the all-participant mode were as follows: for VR, 100% (PM-8), 98.9% (QA), and 90.0% (RBM); for HLSR, 99.6% (RBM), 98.5% (PM-8), and 82.2% (QA); and for HLGR, 99.6% (RBM), 99.3% (PM-8), and 98.1% (QA). The ability of the PM-8 to detect VR and HLAR in enterococci was comparable to those for reference susceptibility and molecular PCR methods and was considered acceptable for routine clinical laboratory use.

Anti-Bacterial Agents

Multicenter comparison of BACTEC 9050 and BACTEC 9240 blood culture systems.

The overall recovery of organisms and time to detection with the BACTEC 9050 and BACTEC 9240 systems were compared in a multicenter evaluation. In the first phase of the study, a total of 4,383 compliant aerobic (Plus Aerobic/F) blood culture sets were processed. There was no significant difference in the recovery of individual groups of organisms with the two systems, with the exception of Streptococcus pneumoniae which was isolated more frequently with BACTEC 9050. False-positive signals occurred more often with BACTEC 9240 (58 cultures) than with BACTEC 9050 (43 cultures), but false-negative cultures were uncommon with both systems (3 cultures for each system). Time to detection of positive cultures of clinically significant organisms was essentially the same with both instruments. In the second phase of the study, 2,431 compliant anaerobic (Plus Anaerobic/F) blood culture sets were processed. There was no significant difference in the recovery of organisms with BACTEC 9050 compared with BACTEC 9240. Significantly (P < 0.03) more false-positive signals occurred with BACTEC 9240 (15 cultures) than with BACTEC 9050 (4 cultures). Likewise, more false-negative cultures occurred with BACTEC 9240 (11 cultures) than with BACTEC 9050 (8 cultures). Time to detection of positive cultures of clinically significant organisms was essentially the same with both systems with the exception of anaerobes (N = 10), which were recovered earlier (P < 0.01) with BACTEC 9240 (35.0 h) than with BACTEC 9050 (61.4 h).

Aerobiosis

Microbial growth inside saline-filled breast implants.

In vitro and in vivo experiments were conducted to determine whether intraluminal saline in breast implants can support the growth of common wound-infecting microorganisms over a prolonged period of time. The bacteria tested were Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Corynebacterium jeikeium, Enterobacter cloacae, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Three fungal species also were tested: Aspergillus fumigatus, Paecilomyces variotii, and Candida albicans. In the in vitro study, four organisms survived in flasks of sterile saline for the 2 weeks in which serial cultures were performed: K. pneumoniae, C. albicans, A. fumigatus, and P. variotii. In the in vivo study, 61 white rabbits (122 implants) received both an experimental implant inoculated with one of the test organisms and a control implant containing only sterile saline. They were sacrificed at 1-, 3-, or 6-month scheduled endpoints. None of the control implants containing sterile saline had positive cultures. In contrast, the intraluminal saline was culture positive for 7 of the 10 inoculated organisms after varying lengths of time: S. epidermidis, E. coli, E. cloacae, K. pneumoniae, P. aeruginosa, A. fumigatus, and P. variotii. Samples of capsular tissue also were cultured. Of the 122 capsular tissue specimens, 21 (17 percent) had positive cultures and surrounded both inoculated and sterile implants. In most instances, capsules that were culture positive contained an organism different from the one that had been inoculated in the group. In only 3 cases was the same organism cultured from both the periprosthetic tissue and the intraluminal saline, and these may represent instances of the inoculated organism migrating through the implants filler valves. The data show that several types of bacteria (particularly gram-negative species) and fungi can grow and reproduce in a restricted saline environment for extended periods of time.

Animals

Controlled clinical laboratory comparison of BACTEC plus aerobic/F resin medium with BacT/Alert aerobic FAN medium for detection of bacteremia and fungemia.

Blood specimens collected from adult patients with suspected sepsis in four medical centers were inoculated into BACTEC Plus/F and BacT/Alert FAN aerobic culture bottles. Both bottles of 7,401 bottle pairs contained the prescribed blood volume of 8 to 12 ml. Bottles were incubated in their respective instruments for a standard 7-day protocol or until the instruments signaled that they were positive. A total of 720 isolates that were judged to represent true infections were recovered from 338 patients; 451 isolates were recovered from both bottles, 143 were recovered from only the Plus/F bottle, and 126 were recovered from only the FAN bottle (P was not significant). Although more Histoplasma capsulatum isolates were recovered from Plus/F bottles (P < 0.005), there were no other statistically significant differences in recovery rates of individual species or groups of organisms between the two systems. Of 329 monomicrobic patient septic episodes, 244 episodes were detected by both blood culture systems, 40 were detected only by the BACTEC system, and 45 were detected only by the BacT/Alert system (P was not significant). There was no significant difference between the two systems in the detection of septic episodes among patients receiving antibiotic therapy at the time of blood cultures. Of the cultures found to be positive within the first 72 h of incubation, detection was on average earlier by the BACTEC system (16.9 h) than by the BacT/Alert system (18.7 h). Larger differences in average time to detection were seen with streptococci (10.7 h by the BACTEC system and 17.9 h by the BacT/Alert system) and yeasts (an average of 29.4 h by the BacT/Alert system versus 37.2 h by the BACTEC system). With the exception of the differences noted above, BACTEC Plus/F aerobic resin and BacT/Alert aerobic FAN blood culture bottles were comparable in their abilities to recover aerobic and facultative organisms.

Adult

Tentative interpretive criteria and quality control parameters for in-vitro susceptibility testing of Neisseria gonorrhoeae to two fluoroquinolones (PD 131628 and grepafloxacin (OPC 17116)).

The susceptibility of Neisseria gonorrhoeae to PD 131628 and grepafloxacin (OPC 17116) was evaluated by agar dilution and disc diffusion methods. A tentative susceptibility category for both fluoroquinolones included strains for which the MICs are < or = 0.06 mg/L and the zones of inhibition are > or = 38 mm for PD 131628 and > or = 37 mm for grepafloxacin. Quality control studies with N. gonorrhoeae ATCC 49226 suggested that agar dilution MIC limits were 0.002-0.008 mg/L and 0.004-0.03 mg/L for PD 131628 and grepafloxacin, respectively. The zone size limits were 50-58 mm for PD 131628 and 44-52 mm for grepafloxacin.

Anti-Infective Agents

Comparison of spiral gradient endpoint and agar dilution methods for susceptibility testing of anaerobic bacteria: a multilaboratory collaborative evaluation.

A multilaboratory collaborative study was carried out to assess the utility of the spiral gradient endpoint (SGE) method for the determination of the antimicrobial susceptibilities of anaerobes and to evaluate the equivalence of the MICs obtained by the SGE method with those obtained by the reference agar dilution method of the National Committee for Clinical Laboratory Standards. The standard deviation of the MIC obtained by the SGE method for the five participating laboratories was +/- 0.26 of a twofold dilution, whereas it was +/- 1 twofold dilution by the reference method. The interlaboratory reproducibility of the results for two control strains tested with imipenem, chloramphenicol, and metronidazole indicated that 96% of the measurements fell within +/- 1 twofold dilution of the mode. The equivalence of the SGE method with the agar dilution method was assessed with a wide variety of anaerobic organisms. The MICs by both methods were within 1 doubling dilution in 93% of the measurements (n = 1,074). Discrepancies generally occurred with those organism-drug combinations that resulted in tailing endpoints (Fusobacterium nucleatum, 86% agreement) or in cases of light growth (Peptostreptococcus spp., 86% agreement).

Agar

Development of interpretive criteria and quality control limits for macrolide and clindamycin susceptibility testing of Streptococcus pneumoniae.

A six-laboratory collaborative study was conducted to develop MIC and zone diameter quality control limits and interpretive criteria for antimicrobial susceptibility testing of Streptococcus pneumoniae with azithromycin, clarithromycin, dirithromycin, and clindamycin. The MICs of all of the agents plus erythromycin for 302 clinical isolates of pneumococci that had been selected with an emphasis on resistant strains were determined by use of the National Committee for Clinical Laboratory Standards (NCCLS)-recommended broth microdilution procedure. The zone diameters of the isolates were also determined for the same agents except erythromycin by the NCCLS disk diffusion test procedure. Repeated testing of S. pneumoniae ATCC 49619 with different sources and lots of media and disks allowed development of MIC and zone diameter quality control ranges for these agents. Interpretive criteria for the MIC of azithromycin were established and were as follows: susceptible, < or = 0.5 microgram/ml; intermediate, 1 microgram/ml; and resistant, > or = 2 micrograms/ml. The interpretive criteria advocated for the MICs of clarithromycin and clindamycin were as follows: susceptible, < or = 0.25 microgram/ml; intermediate, 0.5 microgram/ml; and resistant, > or = 1 microgram/ml. Comparison of MICs and disk diffusion zone diameters led to the development of interpretive criteria for the zone diameters for azithromycin, clarithromycin, and clindamycin that correlated well with these MIC breakpoints. Testing of this organism collection also led to the reestablishment of the erythromycin MIC breakpoints as being identical to those of clarithromycin, which resulted in equivalent cross-susceptibility and cross-resistance for the three macrolides that are currently marketed in the United States. Thus, the susceptibility of pneumococci to azithromycin and clarithromycin can be predicted accurately by testing only erythromycin in clinical laboratories. This recommendation, as well as the interpretive and quality control criteria that are described, have been accepted by NCCLS and are included in the latest NCCLS susceptibility testing guidelines.

Anti-Bacterial Agents

Contemporary testing for enteric pathogens: the potential for cost, time, and health care savings.

We sent a questionnaire to 79 clinical microbiology laboratories seeking information on contemporary practices when investigating for bacterial and protozoan enteric pathogens. Data from the 67 respondents (response rate of 85%) showed that a minority of laboratories (40% for stool culture and 45% for ova and parasite [O&P] examinations) had restrictions for testing in place and that fewer laboratories (24% for stool culture and 19% for O&P examinations) rejected specimens from patients who had been in the hospital for > 3 days. Using two estimates, 15 and 40%, for the proportion of all specimens received from patients in the hospital for > 3 days, we calculated savings for the average hospital in this survey. Reagent savings of $4,000 to $10,000 and time savings of 274 to 731 h per year might have been realized. Moreover, between $26,000 and $71,000 in patient charges could have been prevented. On the basis of this survey, wider application of rejection criteria when testing for enteric pathogens appears possible. If implemented, savings to the nation's health care system could be between $27 and $73 million a year.

Animals

Tests for susceptibility of Streptococcus pneumoniae to cefdinir: proposed interpretive criteria and quality control parameters for both microdilution and disc diffusion methods.

Proposed quality control (QC) parameters for susceptibility testing of Streptococcus pneumoniae to cefdinir were developed following the procedure recommended by the National Committee for Clinical Laboratory Standards. The proposed QC MIC range for microdilution susceptibility testing of S. pneumoniae ATCC 49619 is 0.06-0.25 mg/L. The proposed QC limits for inhibitory zone diameters of S. pneumoniae ATCC 49619 around 5 micrograms cefdinir disks is 26-31 mm. We recommend the following for tentative interpretive criteria for determining the susceptibility of S. pneumoniae to cefdinir: susceptible, MIC < or = 0.5 mg/L or inhibition zone diameter > or = 23 mm; intermediate, MIC 1.0 mg/L or inhibition zone, 20-22 mm; resistant, MIC > or = 2.0 mg/L or inhibition zone diameter, < or = 19 mm for broth microdilution and disc diffusion tests, respectively.

Cefdinir

Paecilomyces variotii contamination in the lumen of a saline-filled breast implant.

This report describes a case of gross contamination with the filamentous fungus P. variotii cultured from an intraluminal saline breast implant removed from a patient 14 months after implantation because of severe capsular contracture. We suspect the fungal contamination occurred when a container of saline was left open in the operating room prior to filling and placement of the implant. This case may be the first documented report of microbial growth and reproduction in the internal environment of a saline implant. We assume that organisms such as P. variotii can survive--and accumulate biomass--on the minute amounts of substrates that diffuse across an implant envelope.

Breast Implants

Multicenter evaluation of the in vitro activity of piperacillin-tazobactam compared with eleven selected beta-lactam antibiotics and ciprofloxacin against more than 42,000 aerobic gram-positive and gram-negative bacteria. In Vitro Susceptibility Surveillance Group.

The in vitro activities of piperacillin-tazobactam, 11 other beta-lactam antibiotics, and ciprofloxacin were evaluated in a large multicenter study. A total of 42,208 organisms, collected from 79 listed medical centers and large community hospitals, were tested using the National Committee for Clinical Laboratory Standards broth microdilution procedure. Imipenem (94.7% inhibition), ciprofloxacin (93.8%), and ceftazidime (90.8%) were the most active antibiotics against 31,426 Gram-negative bacilli. Piperacillin-tazobactam (88.8%) and ticarcillin-clavulanate (86.4%) were the most active combination antibiotics against these organisms. Ampicillin-sulbactam (97.8%), piperacillin-tazobactam (96.4%), and imipenem (96.9%) were the most active antibiotics against 10,782 Gram-positive cocci. Overall, the most active antibiotics against all organisms tested were imipenem (95.2%), ciprofloxacin (91.0%), and piperacillin-tazobactam (90.8%).

Anti-Bacterial Agents

Development of interpretive criteria and quality control limits for broth microdilution and disk diffusion antimicrobial susceptibility testing of Streptococcus pneumoniae.

A five-center collaborative study was undertaken to develop quality control and specific interpretive criteria for susceptibility testing of Streptococcus pneumoniae against 12 antimicrobial agents. MICs were determined for 248 pneumococcal clinical isolates (with an emphasis on resistant strains) by use of the National Committee for Clinical Laboratory Standards (NCCLS)-recommended broth microdilution procedure incorporating lysed horse blood-supplemented Mueller-Hinton broth. NCCLS disk diffusion testing was also performed for each isolate by using Mueller-Hinton sheep blood agar incubated in 5% CO2. Repetitive testing of S. pneumoniae ATCC 49619 with different sources and lots of media and disks allowed development of quality control ranges which encompassed approximately 95% of MIC and zone size values observed in the study. Good intra- and interlaboratory reproducibilities were seen with these testing methods and all of the drugs examined. On the basis of the results of this study, MIC interpretive criteria are proposed for 11 agents. Comparisons of MICs and disk diffusion zone sizes allowed disk diffusion zone size interpretive criteria to be proposed for five drugs and confirmed the use of the oxacillin disk test for prediction of penicillin susceptibility among pneumococci. Excessive numbers of minor-category interpretive errors precludes recommendation at this time of the disk diffusion method for testing of pneumococci against five of the drugs. Use of these proposed quality control and interpretive criteria should provide for reproducible test results and allow recognition of recently emerging resistance among pneumococcal clinical isolates.

Diffusion

Prevalence of Corynebacterium urealyticum in urine specimens collected at a university-affiliated medical center.

Corynebacterium urealyticum (formerly Corynebacterium group D2) has been implicated as a cause of alkaline-encrusted cystitis and urinary tract struvite calculi. Despite preselecting urine specimens with neutral and alkaline pHs and using prolonged incubation on a selective medium, isolation of this organism was rarely observed in a population of hospitalized patients. We do not recommend routine cultures for this organism unless the urine is alkaline and struvite crystals, leukocytes, and erythrocytes are present.

Academic Medical Centers

Laboratory detection of anaerobic bacteremia.

The possibility that mycobacteria may be found in blood cultures is a relatively recent discovery. Immunocompromised patients, especially those with advanced HIV infection, are often mycobacteremic with Mycobacterium tuberculosis or M. avium complex. Laboratory methods provide sensitive detection options for these pathogens.

Bacteremia

In vitro activity of ceftriaxone and other cephalosporins against 602 clinical isolates of staphylococci from geographically diverse medical centers.

The in vitro activity of ceftriaxone and six additional antimicrobial agents (ceftizoxime, cefoperazone, cefuroxime, fleroxacin, ciprofloxacin, and trimethoprim/sulfamethoxazole) was assessed or 602 recent clinical isolates of staphylococci from six geographically distinct medical centers in North America. All seven antimicrobial agents were active (90-100% of strains susceptible) against oxacillin-susceptible (OS) strains of Staphylococcus aureus (OSSA) and coagulase-negative staphylococci (OSCNS) but had limited activity against oxacillin resistant (OR) staphylococci. Our assessment of the in vitro antistaphylococcal activity of ceftriaxone against contemporary isolates of Staphylococcus aureus and coagulase-negative staphylococci indicates that the activity versus OS staphylococci has not changed over the past decade despite widespread use of the drug. It appears that these agents will continue to be useful for empiric therapy in those centers in which OR strains are uncommon.

Academic Medical Centers

In vitro activity of nine fluoroquinolone antibiotics against 200 strains of enterococci.

The in vitro activity of nine fluoroquinolones against 200 recent clinical isolates of enterococci was evaluated. The relative activity was WIN57273 > sparfloxacin > ciprofloxacin, temafloxacin > ofloxacin > fleroxacin, lomefloxacin, norfloxacin > enoxacin. Susceptibility to gentamicin or streptomycin did not influence the activity of the fluoroquinolones.

Anti-Infective Agents

In vitro comparison activity of OPC-17116, a new fluoroquinolone, against more than 5,000 recent clinical isolates from five medical centers.

The in vitro activity of OPC-17116, a new fluoroquinolone, was compared with those of ciprofloxacin, ofloxacin and cefixime. A total of 5,231 fresh clinical isolates from five geographically separate US medical centers were tested. The primary results of this study are: 1. OPC-17116 is very active against Enterobacteriaceae [MIC90s, 0.015-1 microgram/ml, except Providencia rettgeri (MIC90, > 8 micrograms/ml)], 2. among nonenteric Gram-negative bacilli (e.g. Xanthomonas maltophilia, MIC90, 2 micrograms/ml) and Gram-positive cocci (e.g. oxacillin-susceptible staphylococci, MIC90s, 0.12-0.5 microgram/ml) OPC-17116 demonstrated higher activity than ciprofloxacin or ofloxacin, 3. the quinolones were more active than cefixime against most of the species tested, especially against Gram-positive and non-enteric Gram-negative organisms. The results of this study, associated with prior documented favorable qualities, support continued investigation of OPC-17116 for clinical use.

Anti-Infective Agents