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Biomedical subjects

E J Baron

Publications and source records attributed to E J Baron.

At least 19 recordsLinked to original sources

Characterization of Salmonella enterica serotype newport isolated from humans and food animals.

Salmonella enterica serotype Newport isolates resistant to at least nine antimicrobials (including extended-spectrum cephalosporins), known as serotype Newport MDR-AmpC isolates, have been rapidly emerging as pathogens in both animals and humans throughout the United States. Resistance to extended-spectrum cephalosporins is associated with clinical failures, including death, in patients with systemic infections. In this study, 87 Salmonella serotype Newport strains were characterized by pulsed-field gel electrophoresis (PFGE) and antimicrobial susceptibility testing and examined for the presence of class 1 integrons and bla(CMY) genes. Thirty-five PFGE patterns were observed with XbaI, and three of these patterns were indistinguishable among isolates from humans and animals. Fifty-three (60%) Salmonella serotype Newport isolates were identified as serotype Newport MDR-AmpC, including 16 (53%) of 30 human isolates, 27 (93%) of 29 cattle isolates, 7 (70%) of 10 swine isolates, and 3 (30%) of 10 chicken isolates. However, 28 (32%) Salmonella serotype Newport isolates were susceptible to all 16 antimicrobials tested. The bla(CMY) gene was present in all serotype Newport MDR-AmpC isolates. Furthermore, the plasmid-mediated bla(CMY) gene was transferable via conjugation to an Escherichia coli strain. The transconjugant showed the MDR-AmpC resistance profile. Thirty-five (40%) of the isolates possessed class 1 integrons. Sequence analyses of the integrons showed that they contained aadA, which confers resistance to streptomycin, or aadA and dhfr, which confer resistance to trimethoprim-sulfamethoxazole. One integron from a swine isolate contained the sat-1 gene, which encodes resistance to streptothricin, an antimicrobial agent that has never been approved for use in the United States. In conclusion, Salmonella serotype Newport MDR-AmpC was commonly identified among Salmonella serotype Newport isolates recovered from humans and food animals. These findings support the possibility of transmission of this organism to humans through the food chain.

Animals↗

Rapid identification of bacteria and yeast: summary of a National Committee for Clinical Laboratory Standards proposed guideline.

Using colony morphology on selected agars, Gram-stain morphology, and a number of 1-step biochemical or enzymatic tests, skilled microbiologists can identify the species of the majority of isolates seen routinely in a clinical laboratory. These results are often available more quickly than and are as accurate as those derived from conventional methods. The National Committee for Clinical Laboratory Standards has produced a guideline that describes tests that can be used to identify a number of aerobic gram-negative rods and gram-positive cocci, a number of commonly isolated anaerobes, and 3 species of yeast. An overview of the organisms included in the guideline, the tests that identify them, and the situations in which rapid testing is appropriate is presented here.

Bacterial Infections↗

Role of clinical microbiology laboratories in the management and control of infectious diseases and the delivery of health care.

Modern medicine has led to dramatic changes in infectious diseases practice. Vaccination and antibiotic therapy have benefited millions of persons. However, constrained resources now threaten our ability to adequately manage threats of infectious diseases by placing clinical microbiology services and expertise distant from the patient and their infectious diseases physician. Continuing in such a direction threatens quality of laboratory results, timeliness of diagnosis, appropriateness of treatment, effective communication, reduction of health care-associated infections, advances in infectious diseases practice, and training of future practitioners. Microbiology laboratories are the first lines of defense for detection of new antibiotic resistance, outbreaks of foodborne infection, and a possible bioterrorism event. Maintaining high-quality clinical microbiology laboratories on the site of the institution that they serve is the current best approach for managing today's problems of emerging infectious diseases and antimicrobial agent resistance by providing good patient care outcomes that actually save money.

Communicable Disease Control↗

Fecal leukocyte stain has diagnostic value for outpatients but not inpatients.

The methylene blue stain for fecal leukocytes (FL) is widely used as an adjunct to slower but more accurate tests of diarrheal etiology, such as stool culture (SCx) or toxin assays for Clostridium difficile. Prior studies investigating the utility of FL for predicting SCx and C. difficile toxin assay (CDTA) results did not evaluate the importance of inpatient versus outpatient status. We conducted a study of patients who submitted a stool specimen to the Stanford Hospital Microbiology Laboratory between May 1998 and April 1999. The results for stool specimens that were tested by FL and by a confirmatory test (either SCx or CDTA) were used to determine whether the FL method helped to predict the results of these tests. Of 797 stools that were tested by FL method and at least one confirmatory test, 502 stools were tested by CDTA, and 473 stools were cultured. The FL test was 14% sensitive and 90% specific for C. difficile with a diagnostic threshold of one white blood cell/high-power field (WBC/HPF). The overall likelihood ratio (LR) for a positive CDTA was 1.4 with a 95% confidence interval (CI) of 0. 5 to 3.7 (P = 0.5) and was similar among inpatients and outpatients. In contrast, the presence of >/=1 WBC/HPF was 52% sensitive and 88% specific for the 27 positive SCx results and helped to predict a positive SCx result (LR, 4.2; 95% CI, 2.7 to 6.5; P < 0.001). The sensitivity of >/=1 WBC/HPF was 57%, and its predictive value for SCx was higher among outpatients (outpatient LR, 5.0; 95% CI, 2.9 to 8.6; P < 0.001; inpatient LR, 1.9; 95% CI, 0.3 to 10.8; P = 0.5). Among inpatients, only 4 (1.5%) of the 273 SCx results were positive, and the presence of >/=1 WBC/HPF was insensitive (25%) and did not predict a positive SCx (LR, 1.9; 95% CI, 0.3 to 10.8; P = 0.5). When the data were reanalyzed using a diagnostic threshold of five WBC/HPF for FL, the predictive power of the FL method was similar. Thus, FL was of no value in predicting CDTA positivity, nor was it helpful in predicting SCx results for inpatients. Neither SCx nor the FL method should routinely be performed on samples from inpatients. Among outpatients, presence of FLs should suggest a bacterial diarrhea in clinically compatible cases.

Bacterial Proteins↗

Multilaboratory validation of rapid spot tests for identification of Escherichia coli.

To validate the accuracy of rapid tests for identification of Escherichia coli, five laboratories sequentially collected 1,064 fresh, clinically significant strains with core criteria of indole-positive, oxidase-negative, nonspreading organisms on sheep blood agar plates (BAP), having typical gram-negative rod plate morphology, defined as good growth on gram-negative rod-selective media. An algorithm using beta-hemolysis on BAP, lactose reaction on eosin-methylene blue or MacConkey agar, L-pyrrolidonyl-beta-naphthylamide (PYR), and 4-methylumbelliferyl-beta-D-glucuronide (MUG) was evaluated. Identifications using the algorithm were compared to those obtained using commercial kit system identifications. One thousand strains were E. coli and 64 were not E. coli by kit identifications, which were supplemented with conventional biochemical testing of low probability profiles. Of the 1,064 isolates meeting the core criteria, 294 were beta-hemolytic and did not require further testing to be identified as E. coli. None of the 64 non-E. coli strains were hemolytic, although other indole-positive, lactose-negative species were found to be hemolytic when further strains were examined in a follow-up study. Of the remaining strains, 628 were identified as E. coli by a lactose-positive and PYR-negative reaction. For nonhemolytic, lactose-negative E. coli, PYR was not helpful, but a positive MUG reaction identified 65 of 78 isolates as E. coli. The remaining 13 E. coli strains required kit identifications. This scheme for E. coli identification misidentified three non-E. coli strains as E. coli, for an error rate of 0.3%. A total of 13 kit identifications, 657 PYR tests, and 113 MUG tests were needed to identify 1,000 E. coli strains with the algorithm. The use of this rapid system saves laboratory resources, provides timely identifications, and yields rare misidentifications.

Algorithms↗

Methicillin-resistant Staphylococcus aureus outbreak: a consensus panel's definition and management guidelines.

OBJECTIVE: To provide medical personnel with a definition of an outbreak of methicillin-resistant Staphylococcus aureus (MRSA) and guidelines for managing potential outbreaks. PARTICIPANTS: Eighteen panel members were chosen from different specialties, types of institutions, and geographic regions. Representatives from the American Society of Consultant Pharmacists, the American Society of Health-Systems Pharmacists, the Society for Healthcare Epidemiology of America, and the National Association of Directors of Nursing Administration participated. CONSENSUS PROCESS: In preparation for the conference, panel members reviewed the literature and wrote abstracts outlining their personal opinions on the core issues, which were circulated to all participants. During a weekend conference, the panel summarized the reviewed literature, defined an MRSA outbreak, and developed management guidelines. EVIDENCE: Published literature, clinical experience, and expert opinion concerning the emergence and subsequent management of MRSA cases in health care institutions. RESULTS: An outbreak of MRSA was defined as either an increase in the rate of MRSA cases or a clustering of new cases due to the transmission of a single microbial strain in the health care institution. An increased rate of cases can be defined statistically or experientially and includes both infected and colonized patients. A potential outbreak should trigger stepwise, multidisciplinary actions consisting of basic epidemiologic procedures (phase I) to form an initial epidemiologic hypothesis of an outbreak (phase II) followed by a standard epidemiologic workup (phase III) and microbiologic studies (phase IV) to confirm the hypothesis. Mupirocin calcium treatments should be considered to decolonize health care workers during the fourth phase, even before typing is completed. CONCLUSIONS: Until studies can be conducted to delineate the effectiveness of different recommendations, the proposed guidelines may provide a useful starting point that can be adapted to meet an individual institution's specific needs.

Disease Outbreaks↗

Quality perceptions of microbiology services. A survey of infectious diseases specialists.

Opinions about the quality of their primary microbiology laboratory were received from more than 500 practicing infectious diseases specialists by a nationally distributed questionnaire. Approximately 92% of the respondents' primary laboratories were hospital-based. These sophisticated users rated the quality of their microbiology laboratories to be generally high, with bacteriology receiving highest scores and parasitology the lowest scores. Fortunately, the serious problems, such as failing to call a critical result and culture mishandled in the laboratory, were experienced rarely. Laboratories directed by pathologists with specialty microbiology training, PHD microbiologists, and infectious diseases specialists were judged to be of highest quality. American Board of Medical Microbiology certification of the laboratory director was related to higher overall quality perceptions. Whereas physician-customer opinions may not directly measure a laboratory's analytic quality, they are an important performance measure on which laboratories can base quality improvement activities in both service and analytical aspects of performance.

Clinical Laboratory Techniques↗

Lack of agreement among two commercial enzyme-linked immunosorbent antibody assays and a conventional immunofluorescence-based method for detecting islet cell autoantibodies.

Two commercial enzyme-linked immunosorbent assays (ELISAs) for antibodies associated with development of insulin-dependent (type 1) diabetes mellitus (IDDM) were evaluated in conjunction with a conventional indirect immunofluorescent-antibody-islet cell antibody (ICA) test and a radioimmunoprecipitation method for detection of insulin autoantibodies in sera from a selected group of patients. The anti-ICA ELISA was positive for only 1 to 17 serum samples from newly diagnosed IDDM patients but yielded false-positive results with 2 of 6 serum samples containing non-diabetes-related autoantibodies. Although the anti-glutamic acid decarboxylase ELISA did not show positive results for sera with other autoantibodies, it was positive for only 4 of 29 serum samples from recently diagnosed IDDM patients and for 49% of 37 indirect immunofluorescent-antibody-ICA test-positive sera. Until the antibodies associated with the development of diabetes are better characterized, allowing better standards for comparison, it will be difficult to evaluate commercial assays in this field.

Adolescent↗

Infectious disease physicians rate microbiology services and practices.

Recent years have seen increasing emphasis on cost containment and quality improvement in clinical laboratory activities. Modifying those activities to enhance clinical relevance is one strategy that should be satisfying to both laboratory scientists and administrators. This guest commentary describes one approach to quality improvement--the use of user surveys to identify areas for improvement. As an initial attempt to define such areas in clinical diagnostic microbiology, infectious disease specialists, targeted for their particular interest and expertise in microbiology laboratory results, were polled and their responses were analyzed. Some of these data have been presented previously (E. J. Baron, D. P. Francis, and K. M. Peddecord, abstr. C-170, p. 520, in Abstracts of the 94th General Meeting of the American Society for Microbiology, 1994; K. M. Peddecord, E. J. Baron, D. P. Francis, and A. S. Benenson, abstr. C-172, p. 520, in Abstracts of the 94th General Meeting of the American Society for Microbiology, 1994; K. M. Peddecord, E. J. Baron, D. P. Francis, and J. A. Drew, Am. J. Clin. Pathol. 105:58-64, 1996). The discussion includes our recommendations for the use of these survey responses, and their limitations, as stimuli to initiate reexamination of certain microbiology laboratory practices in the interest of developing more cost-effective and clinically relevant protocols.

Bacteremia↗

National survey of the in vitro spectrum of piperacillin-tazobactam tested against more than 40,000 aerobic clinical isolates from 236 medical centers.

Hospital microbiology laboratories from 41 states participated in a bacterial antimicrobial susceptibility study comparing in vitro results generated by the standardized disk diffusion method. Over 41,000 freshly isolated aerobic and facultative strains, representing all specimen types (except stools and urines), were tested for their susceptibility to piperacillin-tazobactam and 21 other antimicrobial agents. Enterococcus spp. was the second or third most common isolate from intraabdominal, gynecologic, and cutaneous infections, confirming its growing importance as a nosocomial pathogen. Escherichia coli was the most frequent isolate overall, despite the exclusion of urinary tract specimens from the study. Pseudomonas aeruginosa was the second most prevalent species, ranking first in frequency of recovery from lower-respiratory-tract specimens. Piperacillin-tazobactam was the most active beta-lactamase inhibitor combination tested against Gram-negative bacteria. Its activity against Gram-positive bacteria and Haemophilus influenzae was similar to that of ampicillin-sulbactam (95-97% susceptible). Imipenem and piperacillin-tazobactam displayed similar spectrums of activity against Gram-positive organisms and Haemophilus influenzae. Against Enterobacteriaceae, piperacillin-tazobactam and ceftazidime exhibited similarly wide spectrums of activity, but with some gaps, particularly among Enterobacter spp. and Citrobacter freundii. In this large-scale in vitro study, piperacillin-tazobactam and imipenem displayed the widest antimicrobial spectrums, inhibiting > 90% of all isolates tested.

Bacteria↗

Survival of anaerobes in original specimens transported by overnight mail services.

Overnight mail delivery was evaluated for its effect on the recovery of facultative and anaerobic microbes in cultures of clinical specimens from patients. Ten clinical specimens, which were collected at different geographic locations and during different weather conditions, were cultured at the site and after overnight delivery to a distant laboratory. Forty-five facultative anaerobic isolates and 48 anaerobes were recovered. There was no significant difference in numbers of strains or relative quantities recovered in cultures of transported and nontransported specimens. With proper collection, transport, and inoculation of specimens, overnight delivery did not compromise recovery of clinically relevant microbes.

Bacteria, Anaerobic↗

"Streptococcus milleri" strains displaying a gliding type of motility.

Isolates belonging to the "Streptococcus milleri" species group that appear to exhibit a gliding type of motility, which is expressed as spreading growth on certain types of agar media, are described. These strains resembled a biotype of "S. milleri" that is usually isolated from genitourinary sources and is notable for its ability to ferment a wide array of carbohydrates. This biotype, which is currently included in the species Streptococcus anginosus, has been implicated in cases of neonatal infection. The "S. milleri" isolates which we studied lacked any observable organelles of motility and gave negative results when they were tested in conventional motility test medium stab cultures. Colonies growing on certain agar media, however, spread over the surfaces of plates and increased in area with increasing time of incubation. Chocolate agar supported maximum spreading, while this characteristic was barely discernible on blood agar. Electron microscopy studies revealed that there was more production of extracellular glycocalyx by motile strains than by a nonmotile isolate having a similar biotype. The results of an analysis of 16S rRNA gene sequences suggested that the motile strains are closely related to S. anginosus and represent a distinct rRNA population within the "S. milleri" species complex.

Base Sequence↗

Genetic aspects of methicillin resistance in Staphylococcus aureus and methods used for its detection in clinical laboratories in the United States.

The mecA gene in methicillin-resistant Staphylococcus aureus (MRSA) directs production of a novel penicillin-binding protein (PBP 2A), an enzyme active in cell wall synthesis. MecA, alone, however, does not determine the degree of resistance expressed by strains of MRSA. Differential resistance or variations in other genes that participate in cell wall synthesis, such as laboratory mutant fem genes, may account, in part, for the heterogeneity of methicillin resistance expression in MRSA. The exact mechanisms of methicillin resistance expression in clinical isolates remain to be elucidated. Laboratories use selective agars containing oxacillin and turbidity pattern recognition programs in automated instruments to identify MRSA, although not all mecA-containing strains are detected. Until a rapid and inexpensive DNA probe assay is widely available, newer test methods such as the E test (AB Biodisk), oxidation-reduction indicators in MIC trays (Alamar), and the rapid fluorescent BBl. Crystal system seem promising.

Anti-Bacterial Agents↗

Comparison of the Accu-CulShure system and a swab placed in a B-D Port-a-Cul tube for specimen collection and transport.

We compared the Accu-CulShure guarded specimen collection device and a swab inserted into a B-D Port-a-Cul transport tube in terms of their efficacy under ideal conditions for recovery of bacteria from 10 decubitus ulcer specimens. Cultures yielded 57 aerobes and 21 anaerobes; 76 isolates were recovered with use of Accu-CulShure, and 72 isolates were recovered with use of Port-a-Cul. Both systems were comparable for recovery of organisms in terms of quantitative and qualitative results.

Adult↗