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F C Tenover

Publications and source records attributed to F C Tenover.

At least 91 records · Page 5Linked to original sources

Three subtypes of the tet(M) gene identified in bacterial isolates from periodontal pockets.

The tet(M) genes were characterized from 84 isolates of 10 different bacterial species isolated from the periodontal pockets of 16 patients with periodontal disease. A 740 bp polymerase chain reaction product from the hypervariable region of the tet(M) structural gene was cleaved with the restriction enzymes AluI and HinfI. Three different restriction patterns were identified for each of the two enzymes. By DNA sequencing, using a direct solid-phase automated sequencing method, the isolates could be grouped into 3 different clusters of tet(M) subtypes. The internal DNA homology within each subtype was 98-100%; the homology between clusters was 89-94%. Two different subtypes were identified in 9 of 10 bacterial species, and the remaining species had 3 different subtypes. One of the subtypes (M3) was seen mainly in the anaerobic isolates. This subtype was different from all earlier sequenced structural tet(M) genes present in the Genbank. Most patients had two different subtypes of tet(M), and a third subtype was seen in the 3 patients who exhibited the greatest variety of tetracycline-resistant bacterial species. It appears that the presence of one subtype of the tet(M) gene within a patient or bacterial species does not prevent the acquisition of another subtype of the same gene. This study identified a new subtype of the tet(M) gene and grouped it into 3 distinct yet highly homologous genetic subtypes.

Bacterial Proteins↗

Tetracycline resistance in Prevotella isolates from periodontally diseased patients is due to the tet(Q) gene.

Tetracycline-resistance in gram-negative periodontal bacteria is often due to the presence of the tet(Q) gene. In the present study the polymerase chain reaction (PCR) was used to examine 54 isolates of gram-negative anaerobic rods (Prevotella intermedia, Prevotella nigrescens and related or Bacteroides-like species) for the presence of the tet(Q) gene. The isolates were recovered from 42 patients with periodontal disease living in northern Europe and North America. An 814 base-pair segment of the tet(Q) gene was amplified from all 41 isolates resistant to tetracycline with minimal inhibitory concentrations of 4 micrograms/ml and above. The presence of the tet(Q) gene was verified using hybridization with a specific oligonucleotide internal to the amplified region and restriction endonuclease digestion with DdeI. A PCR product of the same size was also amplified from one tetracycline susceptible isolate (minimal inhibitory concentration = 0.5 microgram/ml). However, this isolate and the one isolate that was resistant to tetracycline at 4 micrograms/ml showed a weaker signal than the remaining isolates when hybridized with the internal probe. Typing of the PCR products using restriction endonuclease digests with AluI and HpaII revealed two clusters of distinct electrophoresis patterns, indicating that two different subtypes of the tet(Q) gene were present in this material. A control strain containing the tet(Q) gene from Bacteroides thetaiotaomicron had a different electrophoresis pattern for AluI. This study indicated that subtypes of the tet(Q) gene in tetracycline-resistant gram-negative periodontal bacteria exist both within the same patient and within the same species.

Base Sequence↗

Evaluation of commercial methods for determining antimicrobial susceptibility of Streptococcus pneumoniae.

Seven commercial systems for antimicrobial susceptibility testing of Streptococcus pneumoniae were evaluated by using a challenge set of 55 pneumococcal isolates with a variety of resistance phenotypes and genotypes. Overall, the results produced by the Pasco and Etest methods were found to be acceptable for all drugs tested except for trimethoprim-sulfamethoxazole testing by the Etest. The Just One system for penicillin MIC testing was also judged to be acceptable (minor error rate, 5.5%). Although the Sensititre and MicroTech methods both produced 12.7% minor errors with penicillin, the Sensititre method classified penicillin-intermediate strains as resistant or vice versa, while four of MicroTech's errors were among intermediate strains that were classified as susceptible. The MicroMedia (minor error rate, 16.4%) and MicroScan Rapid (minor error rate, 63.6%) methods produced unacceptably high levels of errors when testing penicillin. Minor error rates for cefotaxime and ceftriaxone ranged from a low of 12.7% (Etest and Sensititre) to a high of 28% (MicroMedia). Error rates were low for erythromycin, tetracycline, and chloramphenicol by most methods with the exception of the MicroScan method, which had a high very major error rate for erythromycin (34.6%). For testing of beta-lactam drugs, the Pasco, Etest, and Just One tests for penicillin are the most accurate methods; the Sensititre method also provided acceptable results.

Anti-Bacterial Agents↗

Evaluation of Alamar colorimetric broth microdilution susceptibility testing method for staphylococci and enterococci.

We compared the results of the Alamar broth microdilution susceptibility testing method with the results of the National Committee for Clinical Laboratory Standards reference broth microdilution method for 119 gram-positive organisms. The strains were tested for their susceptibilities to 20 antimicrobial agents. Only appropriate antimicrobial agents were evaluated for each species of bacteria. Absolute categorical agreement between the reference method and the test method was 91.5% for enterococci, 99.8% for oxacillin-susceptible staphylococci, and 97.4% for oxacillin-resistant staphylococci. Essential agreement (percent complete agreement plus percent minor errors) was > 99% for all organisms tested. The results for enterococci showed no very major errors, one major error with ofloxacin, and numerous minor errors with the quinolones. However, all except one of the minor errors were within +/- 1 log2 dilution of the reference result. For staphylococci, only 2 very major errors (one each with chloramphenicol and oxacillin), 1 major error (chloramphenicol), and 15 minor errors (multiple drugs) were observed. The Alamar colorimetric system was easy to use and the results were easy to read. It appears to be an acceptable method for antimicrobial susceptibility testing of staphylococci and enterococci.

Colorimetry↗

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↗

The best of times, the worst of times. The global challenge of antimicrobial resistance.

The development of resistance to antimicrobial agents by many bacterial pathogens has compromised traditional therapeutic regimens, making treatment of infections more difficult and frequently more expensive. Three factors have contributed to the development and spread of resistance: mutations in common genes that extend their spectrum of resistance, transfer of resistance genes among diverse microorganisms and increases in selective pressures in and outside of the hospital environment that enhance the development of resistant organisms. Some new resistance mechanisms are difficult to detect in the laboratory. Thus, resistant microorganisms may go unnoticed until they are widely disseminated in a hospital. The challenge for pharmacists, microbiologists and physicians is not only to contain the spread of existing resistant organisms, but also to prevent the emergence of new resistant pathogens by encouraging the rational and prudent use of antimicrobial agents.

Anti-Bacterial Agents↗

A transferable multiple drug resistance plasmid from Vibrio cholerae O1.

Ten multiple antimicrobial-resistant isolates of Vibrio cholerae O1 isolated from patients in Uganda were characterized, and the transferability of resistance to bacteria of diverse origins was investigated. The isolates were toxigenic and belonged to biotype E1 Tor, serotype Ogawa, and ribotype 8, and possessed a 130-MDa plasmid of incompatibility group 6-C. This plasmid, designated pRVC1, was shown to confer resistance to trimethoprim (mediated by a dhfrI gene), sulfonamides (a suII gene), tetracycline [a tet(C) gene], chloramphenicol (a catI gene), ampicillin (a beta-lactamase gene other than blaTEM or blaSHV), and streptomycin. pRVC1 proved to be transmissible at frequencies between 1 x 10(-1) and 5 x 10(-6) transconjugants per recipient to a variety of bacterial pathogens, including those of humans, animals, and fish. Most efficient transfer was observed from V. cholerae to strains of Shigella flexneri, Escherichia coli, Vibrio parahaemolyticus, and three Aeromonas species. The present in vitro study suggests that pRVC1 may spread from V. cholerae to other bacteria pathogenic to man, animals, and fish in natural environments.

Animals↗

Leptotrichia buccalis bacteremia in patients treated in a single bone marrow transplant unit.

We describe four cases of bacteremia due to Leptotrichia buccalis (an organism that is part of the normal human oral flora) that occurred in a bone marrow transplant unit over a 3-month period. All of the patients were neutropenic, all had mucositis or esophagitis, and all were receiving antimicrobial prophylaxis with ciprofloxacin and vancomycin (drugs to which Leptotrichia is resistant). One patient died of adult respiratory distress syndrome; the others had minimal symptoms. Pulsed field gel electrophoresis of bacterial DNA digested with Sma 1 demonstrated a unique banding pattern for each isolate, indicating that the isolates belonged to distinct strains. Quantitative gas-liquid chromatography of whole-cell free fatty acids confirmed the uniqueness of the strains, obviating the need to search for a common source of infection. We postulate that this outbreak resulted from antibiotic selection pressure on the oral flora in patients who had been compromised by severe neutropenia and mucosal disruption.

Adult↗

Eradication of endemic methicillin-resistant Staphylococcus aureus infections from a neonatal intensive care unit.

To control infections with endemic methicillin-resistant Staphylococcus aureus (MRSA) in a neonatal intensive care unit (NICU), triple dye was applied to the umbilical cords of infants in the intermediate-care but not the intensive-care area. The rate of MRSA infection, adjusted for time and intensity of care, decreased in the intermediate-care area (rate ratio, 0.35; 95% confidence interval [CI], 0.14-0.87; P < .01) but not in the intensive-care area (rate ratio, 0.92; 95% CI, 0.41-2.24; P = .48). After 22 months, the rate increased in both areas (Mantel-Haenszel rate ratio, 1.7; 95% CI, 1.0-2.8; P < .05) after overcrowding and understaffing increased. After temporary reduction of overcrowding and understaffing, extension of triple dye use to the intensive-care area and dedication of an infection control nurse to the NICU, MRSA colonization and infection rates decreased to near zero in both areas (infection rate ratios, 0.09 and 0.11, respectively; P < .005). The endemic MRSA strain, identified by pulsed-field gel electrophoresis, was eradicated.

Birth Weight↗

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↗

Detection of tet(M) and tet(O) using the polymerase chain reaction in bacteria isolated from patients with periodontal disease.

The polymerase chain reaction was used to examine 114 tetracycline-resistant anaerobic and facultative anaerobic bacterial isolates from patients with periodontal disease for the tet(M) and tet(O) genes. A 740-base-pair fragment of the tet(M) gene was amplified from 84 of 114 isolates, and a 519-base-pair fragment of the tet(O) gene was amplified from 13 streptococcal isolates. Six of 7 tetracycline-resistant isolates of Veillonella spp. and tetracycline-resistant isolates of Eubacterium spp. (n = 3), Eubacterium saburreum (n = 1), Streptococcus intermedius (n = 5) and Gemella morbillorum (n = 2) all harbored the tet(M) gene. The tet(M) and tet(O) negative as well as selected positive isolates were tested for the tet(K) and tet(L) genes using DNA probes. All isolates of Staphylococcus spp. (n = 11) hybridized with the tet(K) probe. None of the isolates tested hybridized with the probe for tet(L). This is the first report of the tet(M) gene in the facultative bacterium G. morbillorum and in E. saburreum.

Bacteria, Anaerobic↗

Tetracycline-resistant micro-organisms recovered from patients with refractory periodontal disease.

Tetracycline in combination with scaling and root planing is frequently used to treat refractory periodontal disease. This study examined tetracycline resistance in bacteria recovered from periodontal pockets of patients with refractory periodontitis. Bacterial isolates resistant to 10 micrograms/ml of tetracycline were isolated from plaque samples of 17 patients, of whom 6 had received tetracycline within 8 weeks prior to sampling. Minimal inhibitory concentrations (MICs) of tetracycline and minocycline were determined by agar dilution. In the 6 patients who had received tetracycline, a mean of 22.9% (+/- 38.2) of the total cultivable subgingival flora were resistant to tetracycline, compared with a mean of 7.2% (+/- 8.5) in the untreated group. Although various organisms were isolated, in most patients, the tetracycline-resistant organisms were dominated by Streptococcus spp. Overgrowth of Candida was found in one patient, and of Enterobacteriaceae in another patient, while small numbers of yeast or Staphylococcus spp. were isolated from the plaque samples of 9 others. 3 out of 4 patients who did not respond to tetracycline treatment had a variety of tetracycline-resistant anaerobic Gram-negative rods present. No correlation was found between increased proportions of tetracycline resistance in the whole bacterial sample and the presence of resistant periodontal pathogens.

Adult↗

Identification of multiple clones of extended-spectrum cephalosporin-resistant Streptococcus pneumoniae isolates in the United States.

We characterized 12 isolates of Streptococcus pneumoniae with various levels of susceptibility of penicillin and extended-spectrum cephalosporins by antimicrobial susceptibility patterns, serotypes, ribotypes, chromosomal DNA restriction patterns by pulsed-field gel electrophoresis, multilocus enzyme electrophoresis patterns, penicillin-binding protein (PBP) profiles, and DNA restriction endonuclease cleavage profiles of pbp1a, pbp2x, and pbp2b. Seven cefotaxime-resistant (MIC, > or = 2 micrograms/ml) serotype 23F isolates were related on the basis of ribotyping, pulsed-field gel electrophoresis, and multilocus enzyme electrophoresis, but they had two slightly different PBP patterns: one unique to strains for which the MIC of penicillin is high (4.0 micrograms/ml) and one unique to strains for which the MIC of penicillin is low (0.12 to 1.0 micrograms/ml). The pbp1a and pbp2x fingerprints were identical for the seven isolates; however, the pbp2b fingerprints were different. An eighth serotype 23F isolate with high-level resistance to cephalosporins was not related to the other seven isolates by typing data but was a variant of the widespread, multiresistant serotype 23F Spanish clone. The PBP profiles and fingerprints of pbp1a, pbp2x, and pbp2b were identical to those of the Spanish clone isolate. An additional serotype 6B isolate with high-level resistance to cephalosporins had unique typing profiles and was unrelated to the serotype 23F cephalosporin-resistant isolates but was related on the basis of genetic typing methods to a second serotype 6B isolate that was cephalosporin susceptible. The serotype 6B isolates had different PBP profiles and fingerprints for pbp1a, but the fingerprints for pbp2x and pbp2b were the same.

Aminoacyltransferases↗

Genetic analysis of clinical isolates of Streptococcus pneumoniae with high-level resistance to expanded-spectrum cephalosporins.

Streptococcus pneumoniae CS109 and CS111 were isolated in the United States in 1991 and have high levels of resistance to expanded-spectrum cephalosporins (MICs of 8 and 32 micrograms of cefotaxime per ml, respectively). CS109, but not CS111, also showed high-level resistance to penicillin. As both strains expressed the serotype 23F capsule, were very closely related in overall genotype, and possessed identical or closely related mosaic pbp1a, pbp2x, and pbp2b genes, it is likely that they have arisen from a recent common ancestor. High-level resistance to expanded-spectrum cephalosporins was entirely due to alterations of penicillin-binding proteins (PBPs) 1a and 2x, since a mixture of the cloned pbp1a and pbp2x genes from the resistant strains could transform the susceptible strain R6 to the full level of cephalosporin resistance of the clinical isolates. Both PBP1a and PBP2x of these strains were more resistant to inhibition by cephalosporins than those of typical highly penicillin-resistant isolates. The pbp1a genes of CS109 and CS111 were identical in sequence, and the fourfold difference in their levels of resistance to cephalosporins was due to a Thr-550-->Ala substitution at the residue following the conserved Lys-Ser-Gly motif of PBP2x. This substitution was also the major cause of the 16-fold-lower resistance of CS111 to penicillin. The pbp2x gene of CS111, in an appropriate genetic background, could provide resistance to 16 micrograms of cefotaxime per ml but only to 0.12 microgram of benzylpenicillin per ml. Removal of the codon 550 mutation resulted in a pbp2x gene that provided resistance to 4 microgram of cefotaxime per ml and 4 microgram of benzylpenicillin per ml. The Thr-550-->Ala substitution in CS111 therefore appears to provide increased resistance to expanded-spectrum cephalosporins but a loss of resistance to penicillin.

Amino Acid Sequence↗

Interpretive criteria and quality control parameters for testing bacterial susceptibility to the fluoroquinolone PD131628.

For testing bacterial susceptibility to PD131628, a 5-micrograms disk and the following tentative interpretive criteria may be used: > or = 19 mm for susceptible (MIC, < or = 1.0 micrograms/ml), 16 to 18 mm for intermediate (MIC, 2.0 micrograms/ml), and < or = 15 mm for resistant (MIC, > or = 4.0 micrograms/ml). For standard quality control strains, the following limits are proposed: for Escherichia coli ATCC 25922, zones of 31 to 41 mm or a MIC of 0.002 to 0.016 micrograms/ml; for Pseudomonas aeruginosa ATCC 27853, zones of 26 to 34 mm or a MIC of 0.12 to 0.5 micrograms/ml; for Staphylococcus aureus ATCC 25923, zones of 27 to 33 mm; for Staphylococcus aureus ATCC 29213, a MIC of 0.03 to 0.12 micrograms/ml; and for Enterococcus faecalis ATCC 29212, a MIC of 0.12 to 0.5 micrograms/ml.

Anti-Infective Agents↗

Multilaboratory evaluation of screening methods for detection of high-level aminoglycoside resistance in enterococci. National Committee for Clinical Laboratory Standards Study Group on Enterococci.

Since the early 1970s, the synergistic activity of an aminoglycoside with a cell wall-active agent has been predicted by determining the ability of an enterococcus to grow in the presence of high levels of the aminoglycoside (usually > or = 2,000 micrograms/ml). However, a variety of media and concentrations of aminoglycosides has been used for this screening procedure. In the present study, we sought to optimize the agar dilution, broth microdilution, and disk diffusion tests used to detect high-level gentamicin and streptomycin resistance in enterococci. For dilution tests, brain heart infusion agar or broth gave the best growth and performance. For agar dilution, 500 micrograms of gentamicin per ml, 2,000 micrograms of streptomycin per ml, and an inoculum of 1 x 10(6) CFU/ml were optimal, while for broth microdilution, 500 micrograms of gentamicin per ml, 1,000 micrograms of streptomycin per ml, and an inoculum of 5 x 10(5) CFU/ml were best. Growth of more than one colony in the agar dilution test was determined to be the best indicator of high-level resistance. For disk diffusion, Mueller-Hinton agar, 120-micrograms gentamicin disks, and 300-micrograms streptomycin disks with breakpoints of no zone for resistance and > or = 10 mm for susceptibility gave the best sensitivity and specificity if results for strains with zones of 7 to 9 mm are considered inconclusive, indicating that a broth or agar test should be performed to determine susceptibility or resistance.

Anti-Bacterial Agents↗