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Biomedical subjects

F C Tenover

Publications and source records attributed to F C Tenover.

At least 163 records · Page 9Linked to original sources
PubMed1989

DNA probes for antimicrobial susceptibility testing.

As DNA probes are used more frequently in the clinical laboratory for the detection and identification of pathogens in clinical samples, a means of determining the antimicrobial susceptibility profile of those pathogens will be required. DNA probes directed to specific resistance determinants offer a solution to this problem. Methods of determining the susceptibility of viruses to antiviral agents can also be accomplished with DNA probes. This article explores the advantages and disadvantages of using hybridization methods for susceptibility testing of organisms contained in clinical samples.

DNA Probes↗
PubMed1989

Chlorine dioxide sterilization of implanted right atrial catheters in rabbits.

The disinfection of right atrial catheters in situ using chlorine dioxide was investigated. Catheters were implanted into rabbits (Oryctolagus cuniculi) and colonized by inoculation of Staphylococcus aureus, Sa-80, into the lumen. All of the catheters were colonized and the difference in numbers of bacteria recovered from animals destined for the control and disinfection groups was not significant. Animals were assigned randomly to the control or disinfection group. Treatment consisted of filling the catheter lumen of the disinfection group with chlorine dioxide and of the control group with sterile physiological saline daily for 15 minutes. In addition, both groups received systemic antimicrobial therapy. Cultures of blood withdrawn from the catheters and by venipuncture were negative for five of the nine control group animals after treatment for 5 days. Four control group catheters failed, after from 3 to 21 treatments, without ever achieving negative cultures. All nine animals in the disinfection group had negative cultures after treatment for 5 days. Subsequently, one animal from each group reverted to positive cultures. All nine control group catheters failed during the study, compared to only three disinfection group catheters (p less than 0.01). At necropsy, culture of cardiac blood, thrombi and catheter tubing sections demonstrated colonization of six in the control group and only one in the disinfection group (p less than 0.05). Rabbits tolerated the chlorine dioxide disinfection well and no adverse signs were noted.

Animals↗
PubMed1988

Detection of two different kanamycin resistance genes in naturally occurring isolates of Campylobacter jejuni and Campylobacter coli.

A total of 225 isolates of Campylobacter jejuni and 54 isolates of Campylobacter coli were screened for resistance to kanamycin. Among these, five resistant isolates of C. jejuni and six resistant isolates of C. coli, all with different plasmid patterns, were identified. Each contained at least one plasmid greater than or equal to 41 kilobases in size. The MIC of kanamycin for all 11 strains was determined to be greater than or equal to 256 micrograms/ml by an agar dilution method. In addition, all of the strains exhibited resistance to tetracycline (greater than or equal to 16 micrograms/ml). Eight of the 11 strains transferred the kanamycin resistance phenotype to other Campylobacter strains by conjugation. DNA from 9 of the 11 strains hybridized to a DNA probe specific for the 3'-O-aminoglycoside phosphotransferase type III gene. The remaining two strains also failed to show homology with DNA probes specific for the genes encoding 3'-O-aminoglycoside phosphotransferase types I, II, and III. The novel kanamycin resistance gene was cloned into the vector pBR322 and was expressed in Escherichia coli. Phosphocellulose paper binding assays on sonicates of the E. coli strain carrying the cloned kanamycin determinant demonstrated significant activity against kanamycin, neomycin, and amikacin but not against butirosin, gentamicin, tobramycin, or lividomycin, suggesting that the enzyme is the product of a 3'-O-aminoglycoside phosphotransferase type of aminoglycoside resistance gene.

Animals↗
PubMed1988

Diagnostic deoxyribonucleic acid probes for infectious diseases.

Virtually all microorganisms contain some unique nucleotide sequences which can be the target of deoxyribonucleic acid probes. Probes have been used successfully to identify a wide variety of pathogens from the simple ribonucleic acid-containing polioviruses to the complex filarial worms Brugia malayi. Probe technology offers the clinical laboratory the potential both to extend the types of pathogens that can be readily identified and to reduce significantly the time associated with the identification of fastidious microorganisms. Over a dozen commercially prepared deoxyribonucleic acid probe tests are now available. This article explores the development of deoxyribonucleic acid probe tests and reviews the sensitivity, specificity, and predictive values of many of the diagnostic probes developed during the last several years. Prospects for newer, more sensitive detection systems for the products of hybridization reactions are also reviewed.

DNA Probes↗
PubMed1988

Cloning and sequencing of a gene encoding an aminoglycoside 6'-N-acetyltransferase from an R factor of Citrobacter diversus.

The aacA1 gene, which encodes a 6'-N-acetyltransferase [AAC(6')-I] mediating resistance to kanamycin, tobramycin, and amikacin, was cloned from the Citrobacter diversus R plasmid pBWH100 into the Escherichia coli vector pBR322. The complete nucleotide sequence of the gene and flanking regions was determined. A protein of approximately 21 kilodaltons was identified when the chimeric plasmid encoding the aacA1 gene was introduced into E. coli maxicells. This value is consistent with the size predicted for a protein translated from the open reading frame of the gene.

Acetyltransferases↗
PubMed1988

Nucleotide sequence analysis of tetracycline resistance gene tetO from Streptococcus mutans DL5.

Streptococcus mutans DL5, isolated from the dental plaque of a pig, was resistant to high levels of streptomycin (Sm, 20 mg/ml), erythromycin (Em, 1 mg/ml), and tetracycline (Tc, greater than 100 micrograms/ml), but contained no detectable plasmid DNA. The Smr and Emr determinants were cloned from cellular DNA on the self-replicating 5-kilobase-pair (kbp) EcoRI fragment of pAM beta 1 and the 4.2-kbp cryptic plasmid pVA380-1, respectively, by transformation of Streptococcus sanguis Challis. Helper plasmid cloning, with a Challis host containing pVA380-1, was required to clone the Tcr determinant of strain DL5 on this vector. A single-colony isolate of the original Tcr clone contained a hybrid plasmid, pDL421, composed of 2.6 kbp of vector DNA and 11.4 kbp of S. mutans DNA. Plasmid pDL421 did not hybridize to plasmids containing the streptococcal Tcr determinants tetL, tetM, and tetN. A shortened derivative of this hybrid plasmid, pDL422, missing a 4.9-kbp HincII fragment from the S. mutans DNA but still encoding Tcr, was obtained by subcloning in S. sanguis Challis. The Tcr gene was located in a 1,917-base-pair open reading frame (ORF) corresponding to a 72-kilodalton protein. The ORF exhibited 99.4% sequence identity with the 1,917-base-pair tetO gene from a strain of Campylobacter coli (W. Sougakoff, B. Papadopoulou, P. Nordmann, and P. Courvalin, FEMS Microbiol. Lett. 44:153-160, 1987). A 1.67-kbp NdeI fragment, internal to the ORF from strain DL5, as well as pDL421 hybridized under stringent conditions to DNA from 10 of 10 Tcr strains of C. coli and Campylobacter jejuni from human and animal sources, but not to DNA from Tcs isolates of these two species.

Amino Acid Sequence↗
PubMed1987

Cloning and expression of a tetracycline resistance determinant from Campylobacter jejuni in Escherichia coli.

The tetracycline resistance gene (tet) from the Campylobacter jejuni plasmid pFKT1025 was cloned into both pUC18 and pBR322 and was expressed when the chimeric plasmids were introduced into Escherichia coli. The location of the tet determinant on the chimeric plasmids was determined by BAL 31 deletion mapping within a 2.25-kilobase (kb) RsaI-HincII fragment. A protein of approximately 70 kilodaltons was consistently produced by E. coli maxicells harboring the cloned tet determinant. A 500-base-pair restriction fragment from within the 2.25-kb tet region was shown to hybridize only to DNA from tetracycline-resistant strains of C. jejuni and C. coli, but not to the DNA of organisms known to carry the streptococcal tetM determinant. No homology was noted between the DNA of 10 tetracycline-resistant isolates of campylobacter and the streptococcal tetL, tetM, or tetN determinants when tested under conditions of high stringency. However, homology was noted between a 5.0-kb HincII restriction fragment containing the tetM determinant and two C. jejuni tet R factors under conditions of reduced stringency.

Campylobacter fetus↗
PubMed1987

Naturally occurring auxotrophs of Campylobacter jejuni and Campylobacter coli.

The nutritional requirements for 439 Campylobacter jejuni isolates and 46 Campylobacter coli isolates were determined by using a previously described chemically defined medium, campylobacter defined medium. With this medium, 45% of both human and nonhuman C. jejuni isolates demonstrated auxotrophic requirements. None of the 46 C. coli isolates studied demonstrated requirements for amino acids on campylobacter defined medium. The most common auxotrophic requirement among C. jejuni isolates was for methionine, which was present as a single requirement or in combination with other markers in 21% of human and 28% of nonhuman isolates. There was no correlation between plasmid carriage and auxotype, and a comparison of the Lior serotypes of 472 of the strains showed a correlation only between proline auxotrophs and Lior serotype 11 for strains isolated in the Seattle-King County region.

Amino Acids↗
PubMed1987

Prevalence and characterization of hippurate-negative Campylobacter jejuni in King County, Washington.

A total of 593 strains of thermophilic Campylobacter species were isolated either from humans with diarrhea or from poultry in King County, Washington. Of these strains, 98 (52 hippurate-positive strains and all 46 of the hippurate-negative strains) were selected for further phenotypic characterization and genetic classification. Hippurate hydrolysis, the test typically used to differentiate Campylobacter jejuni and C. coli, did not always correlate with the genetic classification. All hippurate-positive strains were classified as C. jejuni. Of the hippurate-negative strains, 20% were C. jejuni, 78% were C. coli, and 2% were C. laridis. Assuming that the remaining hippurate-positive strains were all C. jejuni, then hippurate-negative C. jejuni represented a small percentage (9 of 556 or 1.6%) of C. jejuni strains but a significant percentage (9 of 46 or 20%) of hippurate-negative strains. This finding suggests that hippurate hydrolysis should not be used as the sole criterion for differentiating thermophilic Campylobacter species, particularly when describing the disease states associated with these organisms.

Animals↗
PubMed1986

Extraintestinal Campylobacter jejuni and Campylobacter coli infections: host factors and strain characteristics.

To determine whether extraintestinal isolates of Campylobacter jejuni and Campylobacter coli are the consequence of unusual host or bacterial characteristics, we studied clinical and bacteriologic features of 24 extraintestinal infections. Common serotypes and auxotypes were present among the extraintestinal isolates. Gastrointestinal isolates were more susceptible to normal human serum than were the systemic isolates; however, the ranges overlapped considerably. Predispositions to systemic spread were present in 52% of patients with extraintestinal infections; isolates from these patients were more often (73%) serum sensitive than were isolates from patients without predispositions (9%; P = .002). By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, no specific protein band was associated with serum resistance, and all isolates of C. jejuni and C. coli had rough-type lipopolysaccharide profiles. Serum susceptibility was inversely correlated with carbohydrate or ketodeoxyoctonate (KDO) fraction of cell weight and directly correlated with KDO:carbohydrate ratio. Our results suggest that either host defects or specific bacterial virulence characteristics, such as serum resistance, possibly related to length of lipopolysaccharide side chain, may be responsible for extraintestinal infections due to C. jejuni and C. coli.

Adolescent↗
PubMed1986

Critical evaluation of the AutoMicrobic system gram-negative identification card for identification of glucose-nonfermenting gram-negative rods.

During a 6-month study we critically evaluated the accuracy of the AutoMicrobic system Gram-Negative Identification Card (Vitek Systems, Inc., Hazelwood, Mo.) in identifying glucose-nonfermenting gram-negative bacilli by testing 419 selected isolates in parallel with a conventional reference method. Of 356 isolates included in the AutoMicrobic system profile, a total of 307 (86.2%) were correctly identified, 36 (10.1%) were not identified, and 13 (3.7%) were misidentified. Fifty-eight of 63 (92%) isolates not included in the profile were correctly reported as "unidentified organisms." Overall, if the first-choice identification was always accepted, only 18 (4.3%) isolates would have been incorrectly reported. When first-choice identifications appended with the special message "questionable biopattern" were rejected, and organisms were screened for characteristic odor and antimicrobial susceptibility before final acceptance of the AutoMicrobic system report, the number of misidentifications was reduced to 5 (1.2%). The average time to identification with the AutoMicrobic system Gram-Negative Identification Card was 15 h. This compares favorably with the 65 h required by the reference method.

Acinetobacter↗
PubMed1986

Comparison of cross-staining reactions by Pseudomonas spp. and fluorescein-labeled polyclonal and monoclonal antibodies directed against Legionella pneumophila.

Commercially prepared polyclonal antisera to Legionella pneumophila are known to cross-react with organisms of the genus Pseudomonas. To determine whether a commercially available monoclonal antibody reagent specific for L. pneumophila would also cross-react with pseudomonads, a two-laboratory study was undertaken to test both monoclonal and polyclonal reagents against 33 isolates of Pseudomonas spp., including 25 Pseudomonas aeruginosa, 4 P. putida, 2 P. maltophilia, 1 P. fluorescens, and 1 P. alcaligenes. Four antisera were tested; polyclonal anti-legionella antisera pools A and B (Centers for Disease Control [CDC], Atlanta, (Ga.), polyclonal 1-6 antisera (BioDx, Inc., Denville, N.J.), and a monoclonal antibody reagent produced by Genetic Systems Corp., Seattle, Wash. All reagents were labeled with fluorescein. Cross-staining reactions were found with the BioDx L. pneumophila antisera and 10 isolates of Pseudomonas. Four of these isolates demonstrated cross-staining with CDC pool A. When tested with individual serotype-specific reagents (CDC), three of four cross-reacted with L. pneumophila serotype 1 antisera; the fourth cross-reacted with serotype 3. No cross-staining reactions were noted with the monoclonal reagent and any of the pseudomonads tested, demonstrating that the Genetic Systems Corp. monoclonal reagent is the most specific of the four reagents tested.

Antibodies, Bacterial↗
PubMed1985

Increased detection of staphylococcal bacteremia using an anti-microbial removal device.

During the period of August 16, 1981, through December 16, 1982, 15-mL blood culture specimens collected at the Seattle Veterans Administration Medical Center (SVAMC) were divided into two aliquots. The first 10-mL aliquot was inoculated directly into aerobic and anaerobic BACTEC (Johnston Laboratories, Towson, MD) vials (DIR); the remaining 5 mL was processed in an resin-containing Antimicrobial Removal Device (Marion Scientific, Kansas City, MO) before transfer to a second, identical set of aerobic and anaerobic vials (ARD). The final volume of inoculated blood from the ARD specimen was half that of the DIR specimen. Both sets of vials were processed using the BACTEC radiometric detection system. One hundred fifty specimen pairs grew 161 significant pathogens; 43 isolates were recovered only from the ARD sample, 39 only from the DIR samples, and 79 from both. Of the 35 isolates recovered from patients receiving anti-microbial agents active against the isolated pathogens, 21 were recovered only from the ARD specimen and 5 only from the DIR specimen (P less than 0.005). Of the 15 S. aureus strains isolated from patients on therapy, 12 were recovered only from the ARD specimen, 2 only from the DIR sample, and 1 from both samples (P less than 0.01). Ten of the 32 isolates of S. aureus recovered from antibiotic-free patients were found only in the ARD specimen and three only in the DIR specimen (P = 0.05). The ARD specimens recovered significantly more S. aureus from all patients regardless of antibiotic status.

Bacteriological Techniques↗
PubMed1985

Campylobacter cinaedi (sp. nov.) and Campylobacter fennelliae (sp. nov.): two new Campylobacter species associated with enteric disease in homosexual men.

In a recent study in Seattle, we isolated Campylobacter-like organisms (CLOs) significantly more often from homosexual men with intestinal symptoms than from asymptomatic heterosexual and homosexual control subjects. Biochemical tests, morphology, and guanine-plus-cytosine content of whole-cell DNA showed that these organisms were similar to, but distinguishable from, other species in the genus Campylobacter. DNA homology tests showed that the Campylobacter-like organisms belong to four unique genetic groups, two of which were phenotypically identical. We developed a rapid screening test to genetically group these organisms and to assess the correlation of each genotype with enteric disease in homosexual men. Three of the four CLO groups were isolated only from men with proctitis, proctocolitis, and/or enteritis. The remaining group, isolated from both symptomatic and asymptomatic homosexual men, was significantly correlated with the presence of polymorphonuclear leukocytes in the rectal secretions of asymptomatic men, a finding that suggests subclinical disease. We have tentatively classified two of the CLO groups into species, Campylobacter cinaedi sp. nov. and Campylobacter fennelliae sp. nov.

Campylobacter↗
PubMed1985

Instability of antibiotic resistance in a strain of Staphylococcus epidermidis isolated from an outbreak of prosthetic valve endocarditis.

Plasmid profiles, phage typing, antibiograms, and biotyping were used to characterize Staphylococcus epidermidis isolated from multiple cultures of blood of four patients with prosthetic valve endocarditis. Epidemiological evidence implicated a common source for these infections. Of 20 clinically significant isolates, 14 exhibited variations from the prototype pattern of multiple resistance to five antibiotics. All isolates tested appeared to be the same strain by phage typing. Of 18 isolates available for plasmid analysis, 10 contained six plasmids of identical size, whereas eight differed from the prototype profile in the loss of one to three plasmids. Loss of resistance to gentamicin, chloramphenicol, erythromycin, and clindamycin but not to methicillin was associated with the loss of specific plasmids. Because antibiotic resistance in this strain of S. epidermidis was unstable, the use of antibiograms alone was not a reliable means of evaluating the relatedness of these multiple isolates.

Anti-Bacterial Agents↗
PubMed1985

Survey of plasmids and resistance factors in Campylobacter jejuni and Campylobacter coli.

A total of 688 isolates of Campylobacter jejuni and Campylobacter coli were screened for the presence of plasmid DNA by agarose gel electrophoresis and were tested for susceptibility to ampicillin, chloramphenicol, erythromycin, streptomycin, and tetracycline. Of the isolates examined, 32% were noted to harbor plasmid DNA, ranging in size from 2.0 to 162 kilobases. Only tetracycline resistance was noted to correlate with the presence of plasmids. Plasmids capable of transferring tetracycline resistance via conjugation ranged in size from 42 to 100 kilobases. The Bg/II and Bc/I restriction endonuclease profiles of 31 plasmids examined showed marked diversity in their banding patterns. Although a high degree of DNA-DNA homology was noted among the Campylobacter spp. plasmids, no homology was noted between these plasmids and tetracycline R factors commonly found in the family Enterobacteriaceae.

Anti-Bacterial Agents↗