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Biomedical subjects

W Witte

Publications and source records attributed to W Witte.

At least 73 records · Page 4Linked to original sources

Discrimination of S. aureus strains by PCR for r-RNA gene spacer size polymorphism and comparison to SmaI macrorestriction patterns.

The size polymorphism of the internal spacer between the 16 S and the 23 S r-RNA genes was studied in S. aureus with the aid of PCR. The patterns of corresponding PCR products were compared with SmaI-generated macrorestriction patterns for definite propagating strains of S. aureus typing phages, for strains with phage pattern 29, phage-group II patterns, phage pattern 94, 96, phage pattern 95 and epidemic methicillin-resistant strains (MRSA). The spacer length polymorphism did not prove to be as discriminative as genomic DNA fragment patterns. However, as shown for S. aureus with phage patterns 29; group II; 94, 96; 95 and also for 4 out of 6 epidemic MRSA, unique patterns of r-RNA gene spacers probably indicate a relatedness among strains which is also suggested by SmaI macrorestriction patterns.

Base Sequence↗

Typing of Staphylococcus aureus by PCR for DNA sequences flanked by transposon Tn916 target region and ribosomal binding site.

The continuous intra- and interhospital spread of multiresistant Staphylococcus aureus demands a rapid molecular typing system. This study describes the fingerprinting of S. aureus by PCR amplification of DNA sequences flanked by the target site for transposon Tn916 and the ribosomal binding site and neighboring nucleotides (target 916-Shine-Dalgarno PCR [tar 916-shida PCR]). Both starting points for PCR are known to be randomly distributed on the S. aureus chromosome. By use of SmaI-macrorestriction patterns as the reference method it was shown that this PCR genotyping discriminates among strains of the major clonal groups of the species S. aureus (strains with phage patterns 29, +, 94, 96, and 95 as well as group II and group III patterns) and identifies the six epidemic methicillin-resistant S. aureus strains prevalent in German hospitals. All of the investigated strains including methicillin-sensitive. S. aureus were typeable. Tar 916-shida patterns are stable during the dissemination of epidemic methicillin-resistant S. aureus among different hospitals.

Bacterial Typing Techniques↗

[MRSA (methicillin-resistant Staphylococcus aureus) infections in patients with pulmonary diseases].

Methicillin-resistant Staphylococcus aureus (MRSA) has become a major nosocomial pathogen. We investigated MRSA-infections in patients with pulmonary diseases referring to epidemiological aspects. Between 9/92 and 2/92 we found MRSA-infections in our hospital in 24 patients (11 female, 13 male, average age 54.6 years). Clinical presentation, main and accompanying disorders and previous antibiotic therapy regimens were registered. Strains were typed using DNA-RFLP and lysotyping. MRSA detection were done in specimen from sputum (12/24) and from the bronchial secret (9/24). In 18/24 cases the MRSA-colonisation was associated with infection. In 15/24 cases the first acquisition of MRSA happened in our hospital, 6/24 times the germ was carried off other institutions and in 3/24 cases it was possibly community acquired. Most frequently patients suffered from bronchial cancer (6/24), from chronical bronchitis (5/24), from pneumonia (4/24) or Cystic fibrosis (4/24). Usually the patients showed other severe comorbidity. 13/24 patients had an antibiotic course before detecting MRSA. Typing revealed a strain already known in different hospitals of Berlin, another known strain of northern Germany and two so far unknown strains. Of interest was a different behaviour of resistance and the lost of resistance of strains in the course. MRSA-infection in pulmonary medicine emerged as a problem mostly in patients with multimorbidity and severe underlying diseases. Change of resistance in strains and new strains were observed.

Adult↗

vanA-mediated high-level glycopeptide resistance in Enterococcus faecium from animal husbandry.

Glycopeptide-resistant Enterococcus faecium strains were isolated from a pig farm and a poultry farm both using avoparcin as a food additive. Such organisms were not isolated in a hen's eggs-producing farm not using avoparcin. Glycopeptide-resistant enterococci were also detected in broiler chicken carcasses that were delivered to a hospital's kitchen. The resistance was determined by the vanA gene as indicated by the detection of the inducible 39-kDa cytoplasmic membrane protein and of a vanA-specific DNA sequence amplified by polymerase chain reaction. Genomic DNA fragment patterns of strains from animal sources were different from each other and also from those of strains isolated in hospitals and from sewage treatment plants. This findings suggest the dissemination of the vanA determinant among different enterococcal strains of distinct ecological origin.

Animal Husbandry↗

Enterococcus faecium strains with vanA-mediated high-level glycopeptide resistance isolated from animal foodstuffs and fecal samples of humans in the community.

The occurrence and the further spread of high-level glycopeptide-resistant, vanA-positive Enterococcus faecium strains outside of hospitals have been investigated. We could isolate such bacteria directly from thawing liquids of commercially produced frozen poultry (chickens, turkeys; no further data on previous feeding with avoparcin were available). In 5 of 13 samples of raw minced meat of pigs originating from 13 different butcher's shops, glycopeptide-resistant E. faecium (VanA type) could be detected after overnight broth cultivation of these samples. No glycopeptide-resistant enterococci could be isolated from meat samples of chickens that were fed without avoparcin. VanA type E. faecium strains were also identified in 12 fecal samples recovered from 100 nonhospitalized humans in the rural area of Saxony-Anhalt federal county. These results suggest a possible role of the food chain in the spread of glycopeptide-resistant E. faecium. Molecular typing (macrorestriction and multilocus enzyme analysis) reveal a wide dissemination of the vanA gene among strains of different ecological origins.

Animals↗

[Analysis and conclusions regarding the epidemic spread of methicillin resistant S. aureus].

Increasing dissemination of Methicillin-resistant S. aureus (MRSA) calls for a more consequent examination of the causal mechanisms. Experiences and results of analysis of three different outbreaks caused by MRSA are described. In summary it is shown that MRSA are especially epidemic virulent and persist over a long range of time in a care unit. Main reasons for these outbreaks obviously were mistakes in hospital hygiene and an inappropriate antibiotic prophylaxis. Predisposed inpatients suffer from severe infections. Interregional dissemination of MRSA observed is mainly due to the transfer of patients between hospitals. Optimization of hospital hygiene especially isolated or cohort nursing of affected patients as well as restriction of antibiotic prophylaxis and therapy are crucial measures against the dissemination of MRSA. For success of these measures information and motivation of staff are necessary.

Antibiotic Prophylaxis↗

Characterization of Staphylococcus intermedius from healthy dogs and cases of superficial pyoderma by DNA restriction endonuclease patterns.

In order to study the possible clonal relation of Staphyloccocus (S.) intermedius from canine superficial pyoderma and from healthy carriers, isolates from pustular swabs and from vaginal, nasal and normal skin sabs were typed using macrorestriction analysis with Sma I and pulsed-field gel electrophoresis. From the size of the resulting fragments the size of the chromosome of S. intermedius could be determined to be roughly 1500 +/- 200 kb on the average. The fingerprints were very heterogeneous though characteristically distinct from patterns of (human) S. aureus as published by others. Strains from superficial pyoderma were not found to be more similar to each other than strains from healthy carriers. Therefore it was concluded that strains from this type of skin infection probably did not belong to a certain subpopulation of S. intermedius, which might have indicated a higher virulence of these strains.

Animals↗

Stability of genomic DNA fragment patterns in methicillin resistant Staphylococcus aureus (MRSA) during the course of intra- and interhospital spread.

The analysis of genomic DNA fragment patterns has revealed as a powerful tool for strain discrimination in Staphylococcus aureus; for use as an epidemiological marker, stability during the course of an outbreak is an essential prerequisite. Genomic DNA fragment patterns (SmaI restriction, pulsed-field electrophoresis) of four different epidemic MRSA strains were compared along with intra- and interhospital and country-wide spread over more than 12 months in Germany. Strain I was isolated from infections in 8 hospitals. In one hospital a subclone arised which differed from the original strain by 4 fragments. Strain II was spread among 4 hospitals, isolates from three of these hospitals exhibited a variability of one to three fragments in the 150-200 kb range. Two hospitals in the Hannover-area were affected by strain III; in 17 isolates of this strain a variability up to three fragments was found in the 170-200 kb range. Strain IV was isolated from 19 cases of infections in 3 hospitals in Berlin. The fragment patterns were completely stable. When S. aureus strains are typed by genomic DNA fragment patterns, a variability in a definite range of molecular masses during the course of an epidemic should be taken into consideration.

Cross Infection↗

Clinical significance and spread of fluoroquinolone resistant uropathogens in hospitalised urological patients.

The minimum inhibitory concentrations (MIC) of ciprofloxacin were determined for 441 uropathogens from patients with complicated and/or hospital acquired urinary tract infections (UTI). None of the Enterobacteriaceae was resistant (MIC > or = 4 mg/l), but 21.7% of enterococci, 28.3% of Pseudomonas spp. and 38.5% of staphylococci were. Subtyping of the strains revealed that with staphylococci there was no clonal spread of resistant strains. In the case of enterococci and Pseudomonas spp., however, cross infections played a major role (46% and 40% respectively). In a retrospective analysis of 370 UTI episodes caused by Pseudomonas aeruginosa (74), enterococci (185) or staphylococci (111) there was no difference between sensitive and resistant strains with respect to clinical aspects and rates of elimination by appropriate anti-bacterial therapy. The rates of spontaneous disappearance without antibacterial therapy ranged from 28% in the case of P. aeruginosa up to 63% in the case of coagulase-negative staphylococci. This implies that especially in UTI caused by gram-positive cocci an indication for antibacterial therapy should be weighted thoroughly and fluoroquinolones should only be used in accordance with sensitivity testing.

Anti-Infective Agents↗

Analysis of nosocomial outbreaks with multiply and methicillin-resistant Staphylococcus aureus (MRSA) in Germany: implications for hospital hygiene.

Two outbreaks of nosocomial infections with MRSA, one in a urological unit in connection with transurethral prostatectomy and the other in an orthopaedic clinic with infections after implantation of prosthetic hips, have been analyzed on the basis of typing MRSA by phage-patterns, plasmid profiles and genomic DNA fragment patterns. Main reasons for these outbreaks were obviously mistakes in hospital hygiene and an inappropriate antibiotic prophylaxis (in the first outbreak a quinolone over about 7 days, in the second a third generation cephalosporin). Both outbreaks could be stopped by measures of hospital hygiene including isolated or cohort nursing of affected patients, and change in antibiotic prophylaxis. Intensive care units (ICUs) are more often affected by MRSA than other clinical settings. As described by the example of an outbreak with MRSA in a municipal hospital, ICUs can play a special role in intrahospital spread of MRSA. The recently observed inter-regional clonal interhospital dissemination of MRSA in Germany is mainly due to a transfer of patients between hospitals; prewarning of the hospital of destination and a number of hygiene measures can prevent further spread.

Anti-Bacterial Agents↗

Characterization of two different clusters of clonally related methicillin-resistant Staphylococcus aureus strains by conventional and molecular typing.

The DNA fragments of 28 distinct isolates of methicillin-resistant Staphylococcus aureus (MRSA) originating from different hospitals in Warsaw and Lodz, were studied. They were obtained by cleavage with restriction endonuclease SmaI and subsequently analysed by pulsed-field electrophoresis. Sixteen different patterns were seen and clusters of related strains were clearly distinguishable. Minor differences in fragment patterns within these clusters and among epidemiologically related strains, revealed genomic rearrangements in the course of clonal dissemination of particular strains. The isolates were also checked for the expression of methicillin resistance. Isolates with heterogenous and homogeneous phenotypes, fell into clearly distinct clusters and thus formed two clonally related MRSA strains. Differences were also seen with phage and biochemical typing, and antimicrobial resistance patterns.

Bacterial Typing Techniques↗

Susceptibility to desferrioxamines and other chelators of coagulase-negative staphylococci.

A total of 233 staphylococci and micrococci belonging to 17 species were tested for their susceptibility to desferrioxamines B, G and E, respectively. Using an agar diffusion method on iron poor media, all of the S. epidermidis, 12 out of 22 S. hominis and 5 out of 22 S. capitis strains were susceptible to desferrioxamines. Among the S. capitis strains tested, two of them were susceptible to desferrioxamine E and resistant to desferrioxamines B and G. All other staphylococci and micrococci tested were resistant to desferrioxamines B, G and E. Different susceptibility to 5 mM ethylenediaminedi-(o-phenylacetic acid) as an artificial chelator demonstrated the existence of additional iron-supplying systems in staphylococci and micrococci.

Chelating Agents↗

Clonal dissemination of two MRSA strains in Germany.

Clonal dissemination of two different MRSA strains, both clumping factor negative, has been observed in Germany for more than a year. Both strains possess the mec-A determinant and each exhibits a characteristic genomic DNA fragment pattern. One strain has spread in the north, the other in the south-west of Germany. Intensive care units are mainly affected by MRSA-infections and probably play a special role in further intra- and inter-hospital spread.

Bacteriophage Typing↗

Cell surface hydrophobicity and adherence to extra-cellular matrix proteins in two collections of methicillin-resistant Staphylococcus aureus.

Non-specific and specific mechanisms of adherence have been examined in two collections of methicillin-resistant Staphylococcus aureus (MRSA). Determination of hydrophobicity by salt aggregation, hydrophobicity indices and of adherence to the extra-cellular matrix proteins fibronectin, vitronectin, laminin and collagen type 1 have failed to reveal any correlation with phage-type, plasmid profile or antibiogram. Further, the strain collections, made over a period of years in two countries, differ markedly in their adherence characteristics; MRSA are heterogeneous in this respect. Such heterogeneity may explain the polarization of views on the epidemicity or 'virulence' of MRSA. With the exception of adherence to collagen a small group of methicillin sensitive S. aureus had characteristics intermediate between the two groups of MRSA.

Agglutination Tests↗

Staphylococcus aureus strains of type 95. Spread of a single clone.

Staphylococcus aureus strains of type 95 in Denmark have increased to a frequency of 20% of the total S. aureus population. A clonal origin and possible subdivision of these strains have been discussed. In the present investigation 35 epidemiologically unrelated S. aureus strains of type 95 as well as reference strains of other types have been analysed by other typing techniques including lectin-typing, multilocus enzyme electrophoresis and pulsed-field gel electrophoresis of genomic restriction fragments. No subdivision could be achieved based on any of these methods and a clonal origin seems therefore possible.

Arsenates↗

[Nosocomial infections with the detection of Staphylococcus aureus in an average hospital--an 11-year analysis].

Nosocomial infections over an 11 years period were monitored prospectively in a district hospital. A total of nosocomial infections among 162.197 patients discharged from 1980-1990 were analyzed. The incidence rate of nosocomial infections was 3.6%. The predominant isolates were Escherichia coli and Staphylococcus aureus (11%). The highest rates of S. aureus infections were established in wound infections and skin and mucosal infections at surgery, gynaecology and paediatrics departments. The number of multiply resistant S. aureus strains has decreased and those of sensitive isolates has increased. 72% of 652 S. aureus isolates have been typed by phage typing and biotyping. Statistical analysis could not establish a significant relationship between phage patterns and nosocomial infections. The spectrum of phage patterns of these nosocomial strains was the same as those of the endogenous colonization of men. Nosocomial outbreaks could not been realized.

Bacteriophage Typing↗