Identification and genetic characterisation of orthopaedic Staphylococcus isolates collected in Italy by automated EcoRI ribotyping

Andollina, Agnese; Cesare, Alessandra De; Bertoni, G.; Modelli, Liliana; Manfreda, Gerardo

doi:10.1016/j.femsle.2004.03.044

Cited by 8 publications

(12 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Ribotyping is based on the analysis of the genomic fragments generated by restriction digestion of the rRNA operons, by highly conserved and present in several copies on the bacterial chromosome. The most conserved regions of the operons allow identification of a bacterium to genus and species level, whereas variable and flanking regions permit discrimination between strains (Andollina et al. 2004).…”

Section: Resultsmentioning

confidence: 99%

OCCURRENCE AND CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS ISOLATED FROM MEAT AND DAIRY PRODUCTS CONSUMED IN TURKEY

Güven

Mutlu

Gülbandılar

et al. 2010

Journal of Food Safety

View full text Add to dashboard Cite

A total of 413 samples of milk, dairy products, meat and meat products were analyzed for the presence of Staphylococcus aureus in a 2‐year survey (2004–2006). One hundred thirty‐eight samples were (33.4%) contaminated with S. aureus. A total of 138 S. aureus strains were isolated and identified. Enterotoksin profiles were determined by Staphylococcal Enterotoxin Test Reversed Passive Latex Agglutination. Their resistance to nine antibiotics were tested. The contamination rate of the meat product samples (48.7%) was significantly higher than milk and dairy products (23.2%). Most of the strains (60.1%) produced staphylococcal enterotoxins. Four strains produced staphylococcal enterotoxin C and staphylococcal enterotoxin D. All of the strains tested were resistant to many of the nine antibiotics. One strain isolated from feta cheese was resistant to all of the antibiotics tested. Penicillin G resistance was the highest (92.7%) among the antibiotics. Seventy‐one strains choosen randomly were compared by the DNA macrorestriction patterns obtained from pulsed field gel electrophoresis following Sma I digestion and 33 pulsotypes were determined. Seventy‐three strains were characterized by automated ribotyping using Eco RI and 8 ribotypes were determined. The same, as well as different pulsotypes, were identified among strains with the identical ribotyping profile. The presence of enterotoxigenic and antimicrobial resistant strains of S. aureus has become remarkably wide spread in the foods examined and poor sanitary conditions during processing may create a health risk for the consumers. The results of genotyping showed high diversity among the strains. PRACTICAL APPLICATIONS These results suggests that the contamination of various food samples by Staphylococcus aureus indicates poor personel hygiene and/or cross contamination in Turkey. Potential of S. aureus strains to produce enteroxins represent a health hazard to the consumers. Prevalence of multiple resistance to the antibiotics of the strains suggests the restricted use of them in animal and human therapy and growth promotion in animals. Genotyping with pulsed field gel electrophoresis (PFGE) revealed more subtypes of S. aureus strains isolated from different food sources than automated ribotyping. Therefore, although both methods could be applied to demonsrate the source and route of the contamination, PFGE should be preferred in future studies of epidemiology of the S. aureus contamination.

show abstract

Section: Resultsmentioning

confidence: 99%

OCCURRENCE AND CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS ISOLATED FROM MEAT AND DAIRY PRODUCTS CONSUMED IN TURKEY

Güven

Mutlu

Gülbandılar

et al. 2010

Journal of Food Safety

View full text Add to dashboard Cite

show abstract

“…RFLP analysis on hsp60 (12) and tuf (9) genes, as well as internal-trancribed spacer PCR followed by direct analysis of amplification products (13) are more simple alternatives, but the former method may be time-consuming and both may reveal intraspecific polymorphisms among S. lugdunensis strains. Automated ribotyping (14), real-time PCR (15), and microarray technology (16) are more sophisticated techniques recruited for coagulase-negative staphylococci speciation, including S. lugdunensis, that have not been extensively evaluated in the literature. This study is the second to report on the development of a speciesspecific single-step PCR protocol for S. lugdunensis identification.…”

Section: Discussionmentioning

confidence: 99%

fbl gene as a species‐specific target for Staphylococcus lugdunensis identification

Chatzigeorgiou

Siafakas

Petinaki

et al. 2010

Clinical Laboratory Analysis

View full text Add to dashboard Cite

Staphylococcus lugdunensis is an unusually virulent coagulase-negative species, associated with severe infections. The present report describes the development of a single-step, species-specific PCR protocol for S. lugdunensis identification. fbl gene, encoding a fibrinogen-binding adhesin, was exploited and assessed as a suitable nucleic acid target. The gene was detected in all 17 S. lugdunensis isolates examined, while no amplification product was obtained from 98 isolates representing 11 staphylococcal and 17 nonstaphylococcal species. Forty-seven percent of the S. lugdunensis strains produced a positive slide coagulase reaction, which is consistent with varying levels of Fbl protein expression within the species.

show abstract

“…However, those techniques remain time-consuming and sometimes of limited value, as for example for CoNS, where commercial identification kits identified only 37% of 177 CoNS isolates with the API 20 Staph system (3). Ribotyping and PCR amplicon sequencing-based methods for identification of CoNS have been described (1,3,5,6,10,14,20,22,25). The methods targeting the sodA or the tuf gene are currently preferred for diagnostic purposes (20,22,25).…”

Section: Discussionmentioning

confidence: 99%

Rapid Identification of Staphylococci Isolated in Clinical Microbiology Laboratories by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

et al. 2007

View full text Add to dashboard Cite

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) of intact bacteria yields a reproducible spectrum depending upon growth conditions, strain, or species. Using whole viable bacteria we describe here the application of MALDI-TOF-MS to the identification of coagulase-negative staphylococci (CoNS). Our aim was, once a bacterium has been recognized as Micrococcaceae, to identify peaks in the spectrum that can be used to identify the species or subspecies. MALDI-TOF-MS was performed using bacteria obtained from one isolated colony. One reference strain for each of the 23 clinically relevant species or subspecies of Micrococcaceae was selected. For each reference strain, the MALDI-TOF-MS profile of 10 colonies obtained from 10 different passages was analyzed. For each strain, only peaks that were conserved in the spectra of all 10 isolated colonies and with a relative intensity above 0.1 were retained, thus leading to a set of 3 to 14 selected peaks per strain. The MALDI-TOF-MS profile of 196 tested strains was then compared with that of the set of selected peaks of each of the 23 reference strains. In all cases the best hit was with the set of peaks of the reference strain belonging to the same species as that of the tested strain, thus demonstrating that the 23 sets of selected peaks can be used as a database for the rapid species identification of CoNS. Similar results were obtained using four different growth conditions. Extending this strategy to other groups of relevant pathogenic bacteria will allow rapid bacterial identification.

show abstract

Identification and genetic characterisation of orthopaedic Staphylococcus isolates collected in Italy by automated EcoRI ribotyping

Cited by 8 publications

References 29 publications

OCCURRENCE AND CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS ISOLATED FROM MEAT AND DAIRY PRODUCTS CONSUMED IN TURKEY

OCCURRENCE AND CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS ISOLATED FROM MEAT AND DAIRY PRODUCTS CONSUMED IN TURKEY

fbl gene as a species‐specific target for Staphylococcus lugdunensis identification

Rapid Identification of Staphylococci Isolated in Clinical Microbiology Laboratories by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

Contact Info

Product

Resources

About