Studies of antimicrobial resistance genes using DNA probes

Tenover, Fred C.

doi:10.1128/aac.29.5.721

Cited by 48 publications

(24 citation statements)

References 45 publications

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“…The number and molecular diversity of resistance determinants in bacteria, although assumed to be vast (34), seem to be limited. Many determinants are related as is shown, again, in this study and also in a similar study on C. difficile (12).…”

Section: Resultsmentioning

confidence: 99%

Homology of a transferable tetracycline resistance determinant of Clostridium difficile with Streptococcus (Enterococcus) faecalis transposon Tn916

Hächler

Kayser

Berger‐Bächi

1987

Antimicrob Agents Chemother

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In several tetracycline-resistant (Tetr) Clostridium difficile strains, homology with the Tn916 part of plasmid pAM120 DNA was observed. This 15-kilobase transposon, carrying a Tetr determinant, was originally found in Streptococcus (Enterococcus) faecalis. Hybridization experiments revealed that at least six of seven Hincll fragments of Tn916, representing >95% of its length, showed homology with DNA of Tetr C. difficile strains. Therefore, a close relationship of the C. difficile Tetr-determining element with the entire Tn9J6 transposon can be assumed, although differences were observed concerning the number of HindlIl cleavage sites within the transposon. In addition to strong hybridization of Tetr determinants of C. difficile with Tn916, weak signals were detected when DNA of Tets C. difficile was hybridized with Tn916. These weak signals could be attributed to a single internal HinclI fragment of Tn916. Clostridium difficile has been described as the most important cause of antimicrobial agent-associated pseudomembranous colitis (2,11,17). Although the pathogenesis of pseudomembranous colitis cannot be explained simply by overgrowth of resistant strains (6), resistance of C. difficile to antibiotics may play a role in the development of the disease. Resistance of C. difficile may also be important from the epidemiological point of view. Nosocomial outbreaks have been reported. In one such study (36), identity of the isolates was claimed and confirmed by bacteriophage typing (H. Hachler and J. Wust, Letter, J. Clin. Microbiol. 20:604, 1984). The responsible strain showed multiple antibiotic resistance (36).Resistance to tetracycline and to other antibiotics has been shown to be transferable between different C. difficile strains (14, 30, 35). Transfer of resistance to tetracycline always occurred independently from that of other resistance markers (35). As far as the mechanism of transfer is concerned only few data are available. The tetracycline resistance (Tetr) determinant is believed to be located on the chromosome and to be transferred by mating procedures only (12,30,35 DNA preparation. Plasmid DNA from C. perfringens or E. coli strains was prepared through alkaline heat treatment by the method of Kieser (16) and either used immediately or purified by CsCl-ethidium bromide equilibrium density gradient centrifugation (20).Chromosomal DNA of C. difficile strains was prepared as follows. Lysis was facilitated by repeated freeze-thawing of pelleted cells and suspension in 50 mM Tris chloride buffer (pH 8), followed by incubation for 15 to 30 min at 37°C with added lysozyme (final concentration, 1 mg/ml). Then EDTA and sodium dodecyl sulfate were added at final concentrations of 1 mM and 1.5%, respectively. After incubation for 15 min in ice, extraction with phenol-chloroform was carried out (20

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Section: Resultsmentioning

confidence: 99%

Homology of a transferable tetracycline resistance determinant of Clostridium difficile with Streptococcus (Enterococcus) faecalis transposon Tn916

Hächler

Kayser

Berger‐Bächi

1987

Antimicrob Agents Chemother

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“…There was an excellent correlation between the resistance phenotypes of the strains studied, as determined by the disk-agar diffusion method or by MICs, and the genotypes inferred from DNA-DNA hybridization experiments using specific probes ( (22).…”

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confidence: 99%

Gene heterogeneity for tetracycline resistance in Staphylococcus spp

Bismuth

Zilhão

Sakamoto

et al. 1990

Antimicrob Agents Chemother

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Nucleotide sequences related to four tet genes were studied by hybridization in 183 clinical Staphylococcus isolates. tet(K) predominated in strains resistant only to tetracycline, while tet(M) was responsible for combined tetracycline and minocycline resistance. In strains harboring both genes, they contributed additively. tet(L) was detected in only five strains, and no hybridization was observed with tet(O).

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“…These include serotyping, biochemical typing, auxotyping, isoenzyme typing, antimicrobial susceptibility testing, bacteriophage typing, bacteriocin typing, and plasmid profiles. Newer typing systems are available that use deoxyribonucleic acid (DNA) probes (188) (including probes derived from chromosomal fragments [196] and ribosomal ribonucleic acid as a probe [69]), polyacrylamide gel electrophoresis of bacterial proteins (136), or chromosomal DNA restriction endonuclease digest patterns (17,20,21,104,107,200). Some of these techniques are being used on organisms other than bacteria (13,25).…”

Section: Introductionmentioning

confidence: 99%

Use of plasmid profiles in epidemiologic surveillance of disease outbreaks and in tracing the transmission of antibiotic resistance

Mayer

1988

Clin Microbiol Rev

152

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Plasmids are circular deoxyribonucleic acid molecules that exist in bacteria, usually independent of the chromosome. The study of plasmids is important to medical microbiology because plasmids can encode genes for antibiotic resistance or virulence factors. Plasmids can also serve as markers of various bacterial strains when a typing system referred to as plasmid profiling, or plasmid fingerprinting is used. In these methods partially purified plasma deoxyribonucleic acid species are separated according to molecular size by agarose gel electrophoresis. In a second procedure, plasmid deoxyribonucleic acid which has been cleaved by restriction endonucleases can be separated by agarose gel electrophoresis and the resulting pattern of fragments can be used to verify the identity of bacterial isolates. Because many species of bacteria contain plasmids, plasmid profile typing has been used to investigate outbreaks of many bacterial diseases and to trace inter- and intra-species spread of antibiotic resistance.

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Studies of antimicrobial resistance genes using DNA probes

Cited by 48 publications

References 45 publications

Homology of a transferable tetracycline resistance determinant of Clostridium difficile with Streptococcus (Enterococcus) faecalis transposon Tn916

Homology of a transferable tetracycline resistance determinant of Clostridium difficile with Streptococcus (Enterococcus) faecalis transposon Tn916

Gene heterogeneity for tetracycline resistance in Staphylococcus spp

Use of plasmid profiles in epidemiologic surveillance of disease outbreaks and in tracing the transmission of antibiotic resistance

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