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G V Doern

Publications and source records attributed to G V Doern.

At least 109 records · Page 6Linked to original sources

Revised disk diffusion interpretive criteria for cefaclor, loracarbef, cefprozil and cefixime when testing Haemophilus influenzae on haemophilus test medium.

The aim of the current five-center collaborative study was to reassess the interpretive criteria for cefaclor, loracarbef, cefprozil and cefixime previously adopted or proposed by the National Committee for Clinical Laboratory Standards (NCCLS) for disk diffusion susceptibility tests with Haemophilus influenzae on Haemophilus Test Medium (HTM) agar. MICs and zones of inhibition were determined using NCCLS methods, HTM and two collections of strains of Haemophilus influenzae. One group of strains consisted of 118 stock organisms taken largely from various recent U.S. antibiotic resistance surveillance studies. The emphasis in this selected group of organisms was on strains that were beta-lactamase negative but ampicillin resistant (BLNAR) by some other mechanism. The second collection of test organisms consisted of 50 recent clinical isolates of Haemophilus influenzae obtained from each of the five participating study centers. This group was considered representative of the type of Haemophilus influenzae currently recovered from clinical sources in the USA. Frequency distribution assessment and error-rate bounded analysis of scattergram comparisons of MICs and zone sizes were used to develop the following zone diameters interpretive for disk diffusion susceptibility tests with Haemophilus influenzae on HTM agar: cefaclor, > or = 20 mm (susceptible, S) and < or = 16 mm (resistant, R); loracarbef, > or = 19 mm (S) and < or = 15 mm (R); and cefprozil, > 18 mm (S) and < 14 mm (R). The respective MIC correlates for all three antimicrobial agents were < or = 8 micrograms/ml (S) and 32 micrograms/ml (R).(ABSTRACT TRUNCATED AT 250 WORDS)

Cefaclor↗

Validation of NCCLS macrolide (azithromycin, clarithromycin, and erythromycin) interpretive criteria for Haemophilus influenzae tested with the Haemophilus test medium. National Committee for Clinical Laboratory Standards.

Some recently marketed macrolide antimicrobial agents possess physiochemical, antimicrobial, and pharmacokinetic advantages that enable their wider clinical use against Haemophilus influenzae infections. A five-laboratory study assessed the validity of existing or proposed azithromycin, clarithromycin, and erythromycin interpretive criteria for tests with H. influenzae isolates. National Committee for Clinical Laboratory Standards (NCCLS) methods, criteria, and quality-control guidelines were used. A total of 350 H. influenzae strains were processed, including fresh clinical isolates (250 strains) and replicate tests of 100 stock cultures sampling strains isolated from 1984 to 91. Azithromycin interpretive criteria (susceptible at < or = 4 micrograms/ml, > or = 12 mm) produced a 99.8% absolute agreement between the minimum inhibitory concentrations and disk diffusion results (0.2% false-susceptible error). Clarithromycin breakpoint criteria (susceptible at < or = 8 micrograms/ml, > or = 13 mm; and resistant at > or = 32 micrograms/ml, < or = 10 mm) produced high minor interpretive error, but < or = 1% combined false-susceptible and false-resistant discrepancies. Erythromycin interpretive guidelines were initially proposed for susceptible at < or = 0.5 microgram/ml, > or = 26 mm. This categorizes nearly all H. influenzae strains as resistant to this older macrolide. The NCCLS should consider the proposed erythromycin criteria for publication in appropriate tables, and a class drug should also be selected (azithromycin) that would best predict macrolide-class susceptibility for those agents indicated by the US Food and Drug Administration for H. influenzae infection chemotherapy (azithromycin and clarithromycin). No serious interpretive problems were observed with the current NCCLS criteria using Haemophilus test medium.

Anti-Bacterial Agents↗

Comparison of two enzyme immunoassays and two latex agglutination assays for detection of cytomegalovirus antibody.

Two commercially available enzyme immunoassays for cytomegalovirus (CMV) immunoglobulin G (IgG), Vidas and Diamedix, and two latex agglutination tests that detect both CMV IgG and IgM, CMV-Scan and CMV-Gen, were evaluated using 165 human serum specimens. Among these, 46.7% contained CMV-reactive antibody. The first three assays had sensitivities of 100%; the sensitivity of the CMV-Gen procedure was 98.7%. The specificities of the four tests were 100, 97.7, 97.3, and 93.2%, respectively.

Antibodies, Viral↗

Optimum recovery of Mycobacterium avium complex from blood specimens of human immunodeficiency virus-positive patients by using small volumes of isolator concentrate inoculated into BACTEC 12B bottles.

Recovery of Mycobacterium avium complex organisms from 412 lysis-centrifugation (Isolator) concentrates of blood specimens obtained from human immunodeficiency virus-positive individuals was attempted with the following media (and Isolator concentrate inoculum volumes): BACTEC 12B broth (0.2 and 1.0 ml), Lowenstein-Jensen slants (0.1 ml), and Middlebrook 7H10/11 agar (0.1 ml). A total of 42 M. avium complex isolates were recovered. The highest rates of recovery and shortest detection times were noted with BACTEC 12B bottles inoculated with 0.2 ml of Isolator concentrate. Middlebrook agar was superior to Lowenstein-Jensen slants. Fluorochrome acid-fast smears performed directly upon Isolator concentrates were of no utility.

Bacteremia↗

Clinical impact of rapid in vitro susceptibility testing and bacterial identification.

During the past decade, a variety of instrument-assisted bacterial identification and antimicrobial susceptibility test systems have been developed which permit provision of test results in a matter of hours rather than days, as has been the case with traditional overnight procedures. These newer rapid techniques are much more expensive than older methods. It has been presumed but not proven that the clinical benefits of rapid testing to patients with infection offset the added cost. The intent of this study was to objectively define the clinical impact of rapid bacterial identification and antimicrobial susceptibility testing. A 1-year study was performed in which infected, hospitalized patients in a tertiary-care, teaching, medical center were randomly assigned to one of two groups: patients for whom identification and susceptibility testing was performed by using a semi-automated, rapid, same-day procedure and those for whom testing was accomplished by using traditional overnight techniques. The two groups were compared with respect to numerous demographic descriptors, and then patients were monitored prospectively through the end of their hospitalization with the aim of determining whether there existed objectively defineable differences in management and outcome between the two groups. The mean lengths of time to provision of susceptibility and identification test results in the rapid test group were 11.3 and 9.6 h, respectively. In the overnight test group, these values were 19.6 and 25.9 h, respectively (P < 0.0005). There were 273 evaluable patients in the first group and 300 in the second group. Other than the length of time required to provide susceptibility and identification test results, no significant differences were noted between the two groups with respect to > 100 demographic descriptors. With regard to measures of outcome, the mean lengths of hospitalization were also the same in both groups. Mortality rates were however, lower in the rapid test group (i.e., 8.8% versus 15.3%). Similarly, statistically significantly fewer laboratory studies, imaging procedures, days of intubation, and days in an intensive or intermediate-care area were observed with patients in the rapid test group. Rapid testing was also associated with significantly shortened lengths of elapsed time prior to alterations in antimicrobial therapy. Lastly, patient costs for hospitalization were significantly lower in the rapid test group. The results of this study indicate the rapid same-day bacterial identification and susceptibility testing in the microbiology laboratory can have a major impact on the care and outcome of hospitalized patients with infection.

Adolescent↗

Manual blood culture systems and the antimicrobial removal device.

In the midst of technologic advances within blood culture microbiology, several manual blood culture systems, which have an important role in the detection of bacteremia, mycobacteremia, and fungemia, are often overlooked. These include traditional broth-based systems, agar-broth biphasic blood culture techniques, a commercial broth-based manual system in which growth is detected by a broth displacement method, and lysis-centrifugation. This article reviews the operational features, advantages, and disadvantages of each.

Anti-Bacterial Agents↗

Comparison of four methods in the diagnosis of Clostridium difficile disease.

Nine hundred forty-five stool specimens from patients suspected of having Clostridium difficile disease were examined using a cell culture cytotoxicity assay (CTA), two enzyme immunoassay (EIA) kits (Cytoclone for toxins A and B; VIDAS for toxin A) and a latex agglutination assay (CDT). One hundred nineteen specimens had positive titers (> or = 90) in the CTA; clinical review of 16 discordant samples and 49 controls supported the significance of 90 as the positive cut-off titer. The performance of the two EIAs and the latex assay was assessed relative to CTA titers of the samples. Sensitivity was < or = 50% for all three assays for the 24 specimens with CTA titers of 90, but it reached 97-100% for the two EIAs and 84% for the latex assay at titers of > or = 2,250. The Cytoclone EIA exhibited higher sensitivity at the lower positive titers. Overall, specificity of the methods ranged from 96.7% (CDT latex assay) to 99.1% (Cytoclone EIA).

Bacterial Proteins↗

Impact of intervention on the frequency with which leaky urine specimens are received in the laboratory.

Prior to intervention, 4.5% of urine specimens received for culturing in the University of Massachusetts Medical Center clinical microbiology laboratory were noted to have leaked during transport. After an extensive educational program was conducted aimed at alerting physician, nursing, and clerical personnel about the problem, a policy was adopted in which physicians were notified by telephone when the laboratory received leaky specimens. The specimens, however, were processed. Leakage rates dropped to 1.7%. A second round of intervention in which leaky urine specimens were not processed was then implemented. No additional decrease in the rate of receipt of leaky urine specimen was noted (that is, 1.9%). One year later, having adopted a policy of not processing leaky urine specimens, the rate of leaky specimens remained at 1.9%. The clinical microbiology laboratory can effectively intervene regarding problems of specimens that have leaked during transport to the laboratory.

Humans↗

In vitro susceptibility test practices with Haemophilus influenzae among College of American Pathologists survey participants in the United States.

Questionnaire results from 5233 clinical microbiology laboratories participating in the College of American Pathologists (CAP) survey program in the United States were used to establish current standards of practice with respect to in vitro susceptibility testing of Haemophilus influenzae. The results of this CAP survey indicated that the recently developed National Committee for Clinical Laboratory Standards (NCCLS) guidelines for H. influenzae susceptibility tests have been widely adopted, particularly with regard to the medium used to perform susceptibility tests. Haemophilus test medium (HTM) is now the most commonly used medium and there exists a general level of satisfaction (approximately 80%) with medium performance. Specific methodologic recommendations of the NCCLS, however, are often not being followed, for example, length and atmosphere of incubation and means of preparing inocula. beta-Lactamase assays constitute a very commonly employed means of assessing ampicillin activity. Among susceptibility test methods, disk diffusion (82.2%) is much more commonly used compared with broth microdilution (17.8%) procedures. Data are provided regarding the most commonly tested antimicrobials as well as some of the problems encounted when using current NCCLS methods for susceptibility tests with H. influenzae. Finally, the CAP survey questionnaire revealed that many laboratories have applied HTM to susceptibility tests with other fastidious bacteria such as pathogenic Neisseria sp., streptococci, and Moraxella catarrhalis.

Culture Media↗

Comparison of MRC-5 and A-549 cells in conventional culture tubes and shell vial assays for the detection of varicella-zoster virus.

A-549 and MRC-5 cells were compared for the detection of varicella-zoster virus (VZV) by conventional tube cell culture and by shell vial assay using fresh specimens. VZV was detected in 33 (32.4%) of 102 specimens by one or all of these methods. In conventional tube culture, seven (21.2%) of 33 were positive in MRC-5 cells and 24 (72.7%) of 33 were positive in A-549 cells, a threefold increase in sensitivity. In shell vial assay, 30 (90.9%) of 33 were positive in MRC-5 cells and 31 (93.9%) of 33 were positive in A-549 cells. The samples tested by A-549 shell vial assay had more positives showing plaque formation. The two shell vial procedures gave the highest levels of sensitivity. A-549 cells provided a more sensitive method of detecting VZV in clinical specimens in both conventional tubes and shell vial assays than did MRC-5 cells.

Cell Line↗

Tsukamurella paurometabolum: a novel pathogen causing catheter-related bacteremia in patients with cancer.

Tsukamurella paurometabolum is a weakly acid-fast, pleomorphic gram-positive bacterium found in soil. Human infection due to this organism has rarely been described, and there are no published accounts of bacteremia. Three cases of bacteremia due to T. paurometabolum and related to long-term use of a central venous catheter in patients with cancer who were receiving chemotherapy are described.

Actinomycetales↗

Optimum use of selective plated media in primary processing of respiratory tract specimens from patients with cystic fibrosis.

A total of 258 respiratory tract specimens from patients with cystic fibrosis were inoculated onto nine different plated media, and the rates of recovery of potential pathogens were compared. Media included sheep blood agar, enriched chocolate agar, MacConkey agar for gram-negative bacilli, chocolate agar containing bacitracin for Haemophilus spp., bromcresol green agar for yeasts, cetrimide agar for Pseudomonas spp., sheep blood agar containing colistin and nalidixic acid for gram-positive cocci, mannitol salt agar for Staphylococcus aureus, and oxidation-fermentation agar containing 300 U of polymyxin B per ml and 2 U of bacitracin per ml (OF-PBL medium) for Pseudomonas cepacia. With two exceptions, all of these media proved useful in recovering potential pathogens from respiratory tract specimens from patients with cystic fibrosis. The two exceptions were cetrimide agar and colistin-nalidixic acid-supplemented sheep blood agar, which were found to be superfluous. In addition, the results of this study further delineated the prevalence of selected bacteria and fungi in respiratory tract secretions from patients with cystic fibrosis. In rank order of frequency of isolation, we recovered isolates of Pseudomonas aeruginosa, Haemophilus parainfluenzae, Candida albicans, S. aureus, Haemophilus influenzae, molds, members of the family Enterobacteriaceae, yeasts other than Candida albicans, miscellaneous gram-negative bacilli, beta-hemolytic streptococci, P. cepacia, and Streptococcus pneumoniae.

Agar↗

Multicenter evaluation of the use of Haemophilus test medium for broth microdilution antimicrobial susceptibility testing of Streptococcus pneumoniae and development of quality control limits.

A five-laboratory collaborative study was undertaken to determine the precision and accuracy of broth microdilution susceptibility tests of Streptococcus pneumoniae isolates performed with Haemophilus test medium (HTM) compared with tests performed with lysed horse blood-supplemented Mueller-Hinton broth (LHB). The intra-and interlaboratory reproducibilities of MICs of 10 antimicrobial agents determined with the two media were found to be quite similar and highly reproducible in both media. On the basis of favorable performance in this study, S. pneumoniae ATCC 49619 is recommended as a quality control strain to assess the performance of HTM when this medium is used for testing of pneumococci. Testing of 293 unique clinical isolates of S. pneumoniae with both media in the respective participant laboratories allowed a direct comparison of MIC results and a calculation of interpretive error rates. Although there were some slight differences between MICs determined with HTM and MICs determined with LHB, few very major or major errors resulted from testing the clinical isolates against the 10 antimicrobial agents. However, MIC-interpretive criteria specific for S. pneumoniae should be developed and promulgated through a national consensus mechanism.

Culture Media↗

Development of revised quality control limits for disk diffusion susceptibility tests of selected cephem antibiotics with Haemophilus influenzae and description of a new control strain.

Inconsistent quality control results in disk diffusion testing of cefaclor, cefamandole, cefonicid, and cefuroxime with Haemophilus influenzae ATCC 49247 and Haemophilus test medium (HTM) prompted a search for an alternative control strain that would provide more reliable results. A five-laboratory study was conducted to evaluate two candidate H. influenzae strains as possible alternatives to the aforementioned strain. Repetitive testing of the candidate strains and H. influenzae ATCC 49247 over several days with a total of six different lots of HTM documented consistent performance of the two candidate strains and confirmed inconsistent results for some of the antibiotics with H. influenzae ATCC 49247. In particular, certain lots of HTM failed to yield cefaclor and cefamandole zone sizes within the quality control range advocated by the National Committee for Clinical Laboratory Standards. Because of the greater consistency offered by the new strains, one was selected (now designated H. influenzae ATCC 49766) to be recommended for routine quality control testing of cefaclor, cefamandole, cefonicid, cefuroxime, and the related carbacephem loracarbef. The new control strain and zone size ranges proposed here have been approved by the National Committee for Clinical Laboratory Standards in place of the previously recommended strain and zone size limits for testing of these five cephem antibiotics.

Ampicillin↗

Laboratory diagnosis of Clostridium difficile-associated gastrointestinal disease: comparison of a monoclonal antibody enzyme immunoassay for toxins A and B with a monoclonal antibody enzyme immunoassay for toxin A only and two cytotoxicity assays.

A total of 320 stool specimens obtained from 262 patients suspected of having Clostridium difficile-associated gastrointestinal disease were examined with two cytotoxicity assays (CTAs) and two commercially available enzyme immunoassays (EIAs). The CTAs were an in-house-developed procedure (University of Massachusetts Medical Center [UMMC], Worcester, Mass.) and a commercial test (Bartels CTA; Baxter Healthcare Corp., West Sacramento, Calif.). One EIA was a monoclonal antibody-based assay for C. difficile toxins A and B (Cambridge Biotech Corp. [CBC], Worcester, Mass.). The other EIA employed monoclonal antibodies directed against only toxin A (Meridian Diagnostics, Cincinnati, Ohio). True-positive and true-negative results were defined on the basis of the results of the four assays, clinical assessments of patients, and the results of other laboratory tests. The sensitivities of the four assays were as follows: Bartels CTA, 100%; UMMC CTA, 97.2%; CBC EIA, 84.5%; and Meridian EIA, 69.0%. The Bartels CTA demonstrated a specificity of 99.2%. The other three assays had a specificity of 100%.

Antibodies, Monoclonal↗

In vitro activity of loracarbef (LY163892), a new oral carbacephem antimicrobial agent, against respiratory isolates of Haemophilus influenzae and Moraxella catarrhalis.

The in vitro activity of a new orally administered carbacephem analog of cefaclor, loracarbef (LY163892), was compared with those of cefaclor and several other oral antimicrobial agents against recent clinical isolates of Haemophilus influenzae and Moraxella catarrhalis. Loracarbef was found to be slightly more active than cefaclor against H. influenzae and had activity essentially equivalent to that of cefaclor for M. catarrhalis. Resistance to loracarbef was uncommon and was noted only with rare beta-lactamase-producing strains of H. influenzae. On the basis of these observations, loracarbef may be of utility in the management of localized, non-life-threatening infections caused by H. influenzae and M. catarrhalis.

Anti-Bacterial Agents↗

Branhamella catarrhalis: phenotypic characteristics.

PURPOSE: This review provides a comprehensive description and discussion of recognized phenotypic characteristics of Branhamella catarrhalis. An emphasis is placed on attributes of this organism that are relevant to its recovery and identification in the clinical microbiology laboratory. In addition, characteristics useful in determining strain identity for use in epidemiologic investigations are addressed. Finally, factors are discussed that may account for the infection-causing potential of B. catarrhalis or at least are of potential consequence to investigations of the pathogenesis of Branhamella disease. CONCLUSIONS: B. catarrhalis is readily isolated from human clinical specimens and can be easily identified using simple, rapid laboratory techniques. Restriction endonuclease analysis of chromosomal deoxyribonucleic acid has proven to be a useful tool in epidemiologic studies. Beta-lactamase isoelectric focusing is of limited value because of the small number of distinct patterns. The lipopolysaccharide and outer membrane proteins of B. catarrhalis have been characterized and found to be relatively non-varying among different strains. Circumstantial evidence exists in support of the hypothesis that the B. catarrhalis beta-lactamase is a virulence determinant.

Humans↗