scholarly journals Detection and Reporting of Organisms Producing Extended-Spectrum β-Lactamases: Survey of Laboratories in Connecticut

1999 ◽  
Vol 37 (12) ◽  
pp. 4065-4070 ◽  
Author(s):  
Fred C. Tenover ◽  
M. Jasmine Mohammed ◽  
Timothy S. Gorton ◽  
Zygmunt F. Dembek
Keyword(s):  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
Test Organism ◽  
National Committee ◽  
Control Strain ◽  
Testing Methods ◽  
Gram Negative ◽  
Extended Spectrum ◽  
Esbl Producers

Extended-spectrum β-lactamases (ESBLs) are enzymes produced in some gram-negative bacilli that mediate resistance to extended-spectrum cephalosporins and aztreonam. They are most common inKlebsiella spp. and Escherichia coli but are present in a variety of Enterobacteriaceae. Resistance mediated by these enzymes can be difficult to detect depending on the antimicrobial agents tested. AmpC β-lactamases are related to the chromosomal enzymes of Enterobacter andCitrobacter spp. and also mediate resistance to extended-spectrum cephalosporins and aztreonam in addition to cephamycins, such as cefoxitin. Unlike ESBLs, however, AmpC β-lactamases are not inhibited by clavulanic acid or other similar compounds. To assess the abilities of various antimicrobial susceptibility testing methods to detect ESBLs, we sent three ESBL-producing organisms, one AmpC-producing organism, and a control strain that was susceptible to extended-spectrum cephalosporins to 38 laboratories in Connecticut for testing. Eight (21.0%) of 38 labs failed to detect extended-spectrum cephalosporin or aztreonam resistance in any of the ESBL- or AmpC-producing isolates. Errors were encountered with both automated and disk diffusion methods. Conversely, seven (18.4%) labs categorized at least some of the four resistant isolates as potential ESBL producers and reported the results with the extended-spectrum cephalosporins and aztreonam as resistant as suggested by current National Committee for Clinical Laboratory Standards (NCCLS) guidelines. The percentage of laboratories that failed to detect resistance in the ESBL or AmpC isolates ranged from 23.7 to 31.6% depending on the type of enzyme present in the test organism. This survey suggests that many laboratories have difficulty detecting resistance in ESBL and AmpC-producing organisms and may be unaware of the NCCLS guidelines on modifying susceptibility testing reports for ESBL-producing strains.

Assessment of Laboratory Performance With Streptococcus pneumoniae Antimicrobial Susceptibility Testing in the United States

1999 ◽  
Vol 123 (4) ◽  
pp. 285-289 ◽  
Author(s):  
Gary V. Doern ◽  
Angela B. Brueggemann ◽  
Michael A. Pfaller ◽  
Ronald N. Jones
Keyword(s):  
United States ◽  
Streptococcus Pneumoniae ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
The United States ◽  
National Committee ◽  
In Vitro Susceptibility ◽  
Susceptibility Tests

Abstract Objective.—To assess the performance of clinical microbiology laboratories in the United States when conducting in vitro susceptibility tests with Streptococcus pneumoniae. Methods.—The results of a nationwide College of American Pathologists Proficiency Survey test sample, in which susceptibility testing of an isolate of S pneumoniae was performed, were assessed with respect to precision and accuracy. Results.—Wide variability was noted among participating laboratories with both minimum inhibitory concentration procedures and disk diffusion susceptibility tests when both methods were applied to S pneumoniae. Despite this high degree of variation, categorical interpretive errors were uncommon. Numerous laboratories reported results for antimicrobial agents that are not recommended by the National Committee for Clinical Laboratory Standards for tests with S pneumoniae. Conclusions.—Current susceptibility testing practices with S pneumoniae in the United States indicate limited precision and a tendency for laboratories to test and report results obtained with antimicrobial agents of questionable therapeutic value against this organism. Continued efforts to standardize susceptibility testing of S pneumoniae in the United States are warranted. In addition, modifications of existing interpretive criteria may be necessary.

scholarly journals Study of the Comparative Activity of Piperacillin/Tazobactam with Currently Available Antibiotics against 8206 Aerobic Isolates

1997 ◽  
Vol 8 (3) ◽  
pp. 147-153 ◽  
Author(s):  
Kevin R Forward ◽  
Donald E Low ◽  
Michel Laverdiere ◽  
Robert Rennie ◽  
Andrew E Simor ◽  
...  
Keyword(s):  
Clinical Laboratory ◽  
National Committee ◽  
Community Hospitals ◽  
Gram Positive ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Extended Spectrum ◽  
Disk Diffusion Testing ◽  
Extended Spectrum Beta Lactamases ◽  
Gram Positive Cocci

OBJECTIVES: To compare the activity of piperacillin-tazobactam with piperacillin and other parenterally administered antibiotics against aerobic Gram-negative bacilli and Gram-positive cocci isolated from across Canada, and to determine the prevalence of resistance mediated by extended-spectrum cephalosporinases.METHODS: Sixty-one laboratories participated. Disk diffusion testing was performed in accordance with methods outlined by the National Committee for Clinical Laboratory Standards. Susceptibilities were performed on 8206 strains.Escherichia coliandKlebsiella pneumoniaewith reduced susceptibilities to third-generation cephalosporins were screened for extended-spectrum beta-lactamases (ESBLs).RESULTS: Piperacillin-tazobactam was active against 92% of the strains, piperacillin against 81% and ticarcillin-clavulanic acid against 88%. Few differences were observed in the relative susceptibility of strains from teaching or community hospitals, from different anatomic sites or from different regions of the country. Aerobic Gram-negative bacilli tested tended to be more susceptible to all the agents than was recently reported in a similar American study. Only 43% ofEnterococcus faeciumwere susceptible to ampicillin and 42% to piperacillin piperacillin with and without tazobactam. Only two enterococcal strains were resistant to vancomycin, and 19 had intermediate zone sizes. Of the 10 strains ofE coliand eight strains ofK pneumoniaewith reduced susceptibility to extended spectrum cephalosporins, only one demonstrated typical ESBL activity.CONCLUSIONS: Canadian aerobic Gram-positive cocci and Gram-negative bacilli remain highly susceptible to many currently available antibiotics. The findings confirm a broad spectrum of activity of piperacillin-tazobactam and indicate that the pattern of susceptibility is quite uniform from different body sites, in both teaching and community hospitals, and across the country.

scholarly journals Evaluation of a Scanner-Assisted Colorimetric MIC Method for Susceptibility Testing of Gram-Negative Fermentative Bacteria

2004 ◽  
Vol 70 (4) ◽  
pp. 2398-2403 ◽  
Author(s):  
Mokhlasur Rahman ◽  
Inger Kühn ◽  
Motiur Rahman ◽  
Barbro Olsson-Liljequist ◽  
Roland Möllby
Keyword(s):  
Bacterial Growth ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
National Committee ◽  
Broth Microdilution ◽  
Gram Negative ◽  
Microdilution Method ◽  
Epidemiological Surveys ◽  
Fermentative Bacteria ◽  
Broth Microdilution Method

ABSTRACT We describe the ScanMIC method, a colorimetric MIC method for susceptibility testing of gram-negative fermentative bacteria. The method is a slight modification of the National Committee for Clinical Laboratory Standards (NCCLS) recommended broth microdilution method that uses a redox indicator 2,3,5-triphenyltetrazolium chloride (TTC) to enhance the estimate of bacterial growth inhibition in a microplate and a flatbed scanner to capture the microplate image. In-house software was developed to transform the microplate image into numerical values based on the amount of bacterial growth and to generate the MICs automatically. The choice of indicator was based on its low toxicity and ease of reading by scanner. We compared the ScanMIC method to the NCCLS recommended broth microdilution method with 197 coliform strains against seven antibacterial agents. The interpretative categorical agreement was obtained in 92.4% of the assays, and the agreement for MIC differences (within ±1 log2 dilution) was obtained in 96% for ScanMIC versus broth microdilution and 97% for a two-step incubation colorimetric broth microdilution versus the broth microdilution method. The method was found to be labor-saving, not to require any initial investment, and to show reliable results. Thus, the ScanMIC method could be useful for epidemiological surveys that include susceptibility testing of bacteria.

scholarly journals Detection of SHV and TEM-type Extended spectrum β-lactamase in bacterial isolates recovered from clinical samples of patients attending military hospitals in South-South Nigeria

2019 ◽  
Vol 38 (3) ◽  
pp. 186
Author(s):  
Helen Oroboghae Ogefere ◽  
Samuel E. Iriah ◽  
Ephraim Ehidiamen Ibadin
Keyword(s):  
Antimicrobial Agents ◽  
Clinical Laboratory ◽  
Clinical Samples ◽  
Bacterial Isolates ◽  
Bacterial Strains ◽  
Gram Negative ◽  
Military Hospitals ◽  
Extended Spectrum ◽  
Phenotypic Method ◽  
Antimicrobial Susceptibility Tests

Background<br />Multi-drug resistant bacterial strains have been increasingly implicated in clinical infections worldwide and beta-lactamase production is one of the commonest mechanisms of resistance in these strains. This study investigated the prevalence of extended spectrum â-lactamase (ESBL)-producing isolates and determined the temoneira (TEM) and sulfhydryl variable (SHV) types implicated in two military hospitals in South-South Nigeria. <br /><br />Methods<br />Three-hundred and eighty (380) consecutive non-duplicate bacterial isolates (Gram negative bacilli) recovered from clinical samples were identified following standard techniques. Antimicrobial susceptibility tests were performed for each isolate following the Clinical Laboratory Standards Institute guidelines. Bacterial isolates recovered which comprised Escherichia coli, Klebsiella spp, Proteus spp and Pseudomonas aeruginosa were screened for ESBL using a phenotypic method (double disc synergy test). All positive isolates were screened for TEM and SHV genes by PCR method. <br /><br />Results<br />Sixty-five isolates (17.1%) were ESBL producing using phenotypic method, E. coli showed the highest ESBL prevalence (24.3%). One isolate was SHV positive (1.5%), 8 (12.3%) were TEM positive while 3 (4.6%) isolates harbored both SHV and TEM genes. Fluoroquinolone - ofloxacin showed marked activity against ESBL-producing isolates (90.8%) while the least active were ceftriaxone (9.2%), ceftazidime (3.1%) and ampicillin (1.5%). <br /><br />Conclusion<br />This study demonstrated that 17.1% of Gram-negative bacilli were ESBL producers. Screening of clinical isolates for ESBL should be implemented. The findings of this study suggest the need for caution in the use of antimicrobial agents in order to curb the incidence of antimicrobial resistance.

scholarly journals Evaluation of the Dade MicroScan MICroSTREP Antimicrobial Susceptibility Testing Panel with Selected Streptococcus pneumoniae Challenge Strains and Recent Clinical Isolates

1998 ◽  
Vol 36 (3) ◽  
pp. 788-791 ◽  
Author(s):  
J. H. Jorgensen ◽  
M. L. McElmeel ◽  
S. A. Crawford
Keyword(s):  
Streptococcus Pneumoniae ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
Reference Panel ◽  
Clinical Isolates ◽  
National Committee ◽  
Broth Microdilution ◽  
Drug Concentrations ◽  
Broth Microdilution Method

The MicroScan MICroSTREP panel is a recently marketed frozen broth microdilution panel for susceptibility testing of various streptococci, including Streptococcus pneumoniae. The panel contains 10 antimicrobial agents in cation-adjusted Mueller-Hinton broth supplemented with 3% lysed horse blood, similar in concept to the National Committee for Clinical Laboratory Standards (NCCLS) reference broth microdilution method for testing streptococci. A group of 210 isolates of S. pneumoniae were selected to include isolates with previously documented resistance to agents incorporated in the MICroSTREP panel, as well as recent invasive clinical isolates. All isolates were tested simultaneously with the MICroSTREP panel and an NCCLS reference panel whose drug concentrations were prepared to coincide with those of the MICroSTREP panel. Of the 210 isolates, 5 failed to grow in the MICroSTREP panel; 3 of those also did not grow in the reference panel. Essential agreement of MICs determined by the two methods (test MIC ± one dilution of the reference MIC) was 99.6% overall and ranged from 98.0% with chloramphenicol to 100% with penicillin, ceftriaxone, erythromycin, tetracycline, and vancomycin. There were no very major or major interpretive category errors resulting from the MICroSTREP panel tests. Minor interpretive category errors ranged from 12.2% with cefotaxime and 9.8% with ceftriaxone (due mainly to clustering of MICs for the selected strains near the breakpoints) to 0% with chloramphenicol and vancomycin. These results indicate that the MicroScan MICroSTREP frozen panels provide susceptibility results with pneumococci that are essentially equivalent to results derived by the NCCLS reference broth microdilution procedure.

scholarly journals Extended-Spectrum β-Lactamases in Ireland, Including a Novel Enzyme, TEM-102

2003 ◽  
Vol 47 (8) ◽  
pp. 2572-2578 ◽  
Author(s):  
Dearbháile Morris ◽  
Colette O'Hare ◽  
Maura Glennon ◽  
Majella Maher ◽  
Geraldine Corbett-Feeney ◽  
...  
Keyword(s):  
Molecular Typing ◽  
Clinical Laboratory ◽  
Enterobacter Cloacae ◽  
Disk Diffusion ◽  
National Committee ◽  
Esbl Production ◽  
Esbl Gene ◽  
Extended Spectrum ◽  
The Family ◽  
Esbl Producers

ABSTRACT Organisms producing extended-spectrum β-lactamases (ESBLs) have been reported in many countries, but there is no information on the prevalence of ESBL-producing members of the family Enterobacteriaceae in Ireland. A total of 925 isolates of ampicillin-resistant members of the Enterobacteriaceae were received from six hospitals in Ireland over a 3-year period from September 1996 to September 1999. Isolates were screened for ESBL production by the double-disk diffusion (DDD) method. DDD-positive isolates that were (i) confirmed as ESBL producers by National Committee for Clinical Laboratory Standards (NCCLS) confirmatory testing and (ii) susceptible to cefoxitin by disk diffusion were considered ESBL producers. By these criteria, 27 (3%) of the ampicillin-resistant members of the Enterobacteriaceae studied were categorized as ESBL producers. Molecular typing suggested that some intra- and interhospital spread of ESBL-producing isolates had occurred. DNA sequencing of amplified bla TEM and bla SHV genes resulted in the detection of a novel bla TEM ESBL gene, bla TEM-102 in two isolates (Klebsiella pneumoniae and Enterobacter cloacae) received from the same hospital but isolated from different patients. The study suggests dissemination of ESBL-producing bacteria within the health care system in Ireland and emphasizes the need for measures to control such spread.

scholarly journals Comparison of Disc Diffusion Methods for the Detection of Extended-Spectrum Beta Lactamase-Producing Enterobacteriaceae

2011 ◽  
Vol 3 (01) ◽  
pp. 033-036 ◽  
Author(s):  
Ravi S Giriyapur ◽  
Namratha W Nandihal ◽  
Krishna B V S. ◽  
Asha B Patil ◽  
Chandrasekhar M R.
Keyword(s):  
Clinical Laboratory ◽  
Confirmatory Test ◽  
National Committee ◽  
P Value ◽  
Close Monitoring ◽  
Accurate Identification ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Esbl Producers

ABSTRACTBackground: Resistance to broad-spectrum β lactams, mediated by extended-spectrum β lactamases (ESBLs), is an increasing problem world wide. This resistance poses problems for in vitro testing and reporting. Increased prevalence of ESBLs among Enterobacteriaceae creates a great need for laboratory testing methods that will accurately identify their presence.Materials and Methods: During the study, the Enterobacteriaceae isolated were tested for the presence of ESBL by the National Committee for Clinical Laboratory Standards (NCCLS) screening test, Jarlier double disc synergy (approximation) test (DDST) and NCCLS phenotypic confirmatory test (PCT), and compared their efficiency in detection.Results: A total of 313 Enterobacteriaceae were isolated and tested for the presence of ESBL. NCCLS PCT identified 200 (63.89%) as ESBL producers and DDST identified 176 (56.23%), with a P-value of <0.001. Among the screening agents, ceftazidime had a better sensitivity (89.49%) and specificity (95.74%).Conclusions: Close monitoring of the susceptibility pattern of isolates and careful spacing with specific discs can identify many ESBL producers. Ceftazidime has a better sensitivity and specificity as a screening agent. A combination of different tests can be useful for accurate identification.

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