Intercenter reproducibility of binary typing for Staphylococcus aureus

Leeuwen, Willem B. van; Snoeijers, S.S.; Werken-Libregts, Christel van der; Tuip, Anita; Zee, Anneke van der; Egberink, Diane; Proost, Monique de; Bik, Elisabeth M.; Lunter, Bjorn; Kluytmans, Jan; Gits, Etty; Duyn, Inge van; Heck, Max; Zwaluw, Kim van der; Wannet, W J B; Noordhoek, G. T.; Mulder, S.; Renders, Nicole; Boers, Miranda; Zaat, S. A. J.; Riet, Daniëlle van der; Kooistra, Mirjam; Talens, Adriaan; Dijkshoorn, Lenie; Reyden, Tanny van der; Veenendaal, Dick; Bakker, Nancy; Cookson, B.; Lynch, Alisson; Witte, Wolfgang; Cuny, Cécile; Blanc, Dominique; Vernez, Isabelle; Hryniewicz, Waleria; Fiett, Janusz; Struelens, Marc; Deplano, Ariane; Landegent, J. E.; Verbrugh, Henri A.; Belkum, Alex van

doi:10.1016/s0167-7012(02)00049-0

Cited by 20 publications

(28 citation statements)

References 22 publications

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“…aureus typing is a useful adjunct in several clinical settings, in addition to its use during dramatic acute outbreaks. Despite the use of several phenotypic and genotypic methods (antibiotyping, phage typing, multilocus enzyme electrophoresis, restriction analysis of cellular DNA, analysis of PCR products, and multilocus sequence typing) (10,13,22,24,31,32, 35,36), indistinguishable or closely related isolates have been detected not only among those responsible for outbreaks, but also among those isolated in different countries, at time intervals of several years, and without any obvious epidemiological links. Indeed, Oliveira et al (27) The aim of the present study was to design a DNA macroarray with several intragenic PCR amplicons to identify S. aureus at the species level and to type S. aureus isolates.…”

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DNA Macroarray for Identification and Typing of Staphylococcus aureus Isolates

et al. 2004

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A DNA macroarray containing 465 intragenic amplicons was designed to identify Staphylococcus aureus at the species level and to type S. aureus isolates. The genes selected included those encoding (i) S. aureus-specific proteins, (ii) staphylococcal and enterococcal proteins mediating antibiotic resistance and factors involved in their expression, (iii) putative virulence proteins and factors controlling their expression, and (iv) proteins produced by mobile elements. The macroarray was hybridized with the cellular DNAs of 80 S. aureus clinical isolates that were previously typed by analyses of their antibiograms and SmaI patterns. The set selected contained unrelated, endemic, and outbreak-related isolates belonging to 45 SmaI genotypes. In a gene content dendrogram, the 80 isolates were distributed into 52 clusters. The outbreak-related isolates were linked in the same or a closely related cluster(s). Clustering based on gene content provided a better discrimination than SmaI pattern analysis for the tested mecA ؉ isolates that were endemic to Europe. All of the antibiotic resistance genes detected could be correlated with their corresponding phenotypes, except for one isolate which carried a mecA gene without being resistant. The 16 isolates responsible for bone infections were distinguishable from the 12 isolates from uninfected nasal carriers by a significantly higher prevalence of the sdrD gene coding for a putative SD (serine-aspartate) adhesin (in 15 and 7 isolates, respectively). In conclusion, the macroarray designed for this study offers an attractive and rapid typing method which has the advantage of providing additional information concerning the gene content of the isolate of interest.The best known staphylococcal species is Staphylococcus aureus, by virtue of its frequent and highly versatile pathogenicity in humans and animals. Isolates belonging to this species are responsible for suppurative infections and syndromes provoked by toxins. Excluding pathologies caused by toxins such as enterotoxins and exfoliative or toxic shock syndrome toxins (20), the pathology of a staphylococcal infection is attributable not to a single factor but to the coordinated actions of several factors whose expression is controlled by several regulatory systems (3,26,29,30). S. aureus is one of the most common causes of nosocomial infections. The emergence of such infections is of particular concern since most isolates, such as methicillin-resistant S. aureus (MRSA), are resistant to several antibiotics (4, 28) and because the spread of these strains in hospitals often increases the overall incidence of nosocomial S. aureus infections in the institution. MRSA clinical isolates with decreased susceptibilities to glycopeptides (1, 17) threaten to compromise our ability to treat hospital-acquired S. aureus infections.S. aureus typing is a useful adjunct in several clinical settings, in addition to its use during dramatic acute outbreaks. Despite the use of several phenotypic and genotypic methods (antibiotyping, phage typi...

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DNA Macroarray for Identification and Typing of Staphylococcus aureus Isolates

et al. 2004

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“…Several alternative typing methods have been published recently in order to replace PFGE. These include multilocus sequence typing (12,22), MecA-associated variable element genotyping (4,15), fluorescent amplified fragment length polymorphism (5), binary typing (23), variable number of tandem repeat analysis (14), and others (1,2,13). In our laboratory we developed multienzyme amplified fragment length polymorphism (ME-AFLP) for typing Klebsiella spp.…”

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Recognition of SCC mec Types According to Typing Pattern Determined by Multienzyme Multiplex PCR-Amplified Fragment Length Polymorphism Analysis of Methicillin-Resistant Staphylococcus aureus

et al. 2005

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Multienzyme multiplex PCR-amplified fragment length polymorphism (ME-AFLP) typing is a reliable and simple method for typing of bacterial species. In this study we analyzed two well-documented strain collections of Staphylococcus aureus and compared ME-AFLP typing results with results of various other typing methods. The discriminatory power of ME-AFLP was found comparable to pulsed-field gel electrophoresis, and typing results were highly concordant. ME-AFLP typing presents a suitable method for prescreening of large strain collections. Furthermore, the obtained typing patterns were found to cluster according to the staphylococcal cassette chromosome mec types of the strains.Staphylococcus aureus is the most significant pathogen involved in nosocomial infections. A rapid epidemiological typing system is needed for the identification of outbreaks and to monitor spread. Some methicillin-resistant S. aureus (MRSA) strains are known to spread more easily and are designated epidemic MRSA, while other strains lack this capacity and are usually associated with sporadic cases. In The Netherlands strict rules for containment of MRSA-infected patients are followed. Due to the policy of "search and destroy," MRSA is not endemic in The Netherlands (24). Pulsed-field gel electrophoresis (PFGE) is regarded as the gold standard for epidemiological typing of MRSA, and a new harmonized protocol for PFGE has been developed enabling the establishment of a European web-based database for comparison of intercenter MRSA PFGE patterns (10). However, PFGE is a time-consuming and tedious method and therefore is not best suited for use in the hospital clinical microbiology laboratory and in large-scale studies of outbreaks. Several alternative typing methods have been published recently in order to replace PFGE. These include multilocus sequence typing (12, 22), MecA-associated variable element genotyping (4, 15), fluorescent amplified fragment length polymorphism (5), binary typing (23), variable number of tandem repeat analysis (14), and others (1, 2, 13). In our laboratory we developed multienzyme amplified fragment length polymorphism (ME-AFLP) for typing Klebsiella spp. (20) and various other gram-negative bacteria. This method relies on the amplification of selections of restriction fragments which represent a total chromosomal image. Since this broadly applicable method would also be very convenient for typing of MRSA we investigated the performance of ME-AFLP typing of MRSA in comparison with PFGE. Our aim was to be able to use ME-AFLP typing for prescreening MRSA strains and thus limit the number of strains to be subjected to PFGE. We used the two well-documented strain collections (17, 18, 19) that were described previously with various different other pheno-and genotyping methods. MATERIALS AND METHODSBacterial strains. The present study includes the use of two well-defined MRSA strain collections. The first set consists of 47 isolates which have been described in great detail (17)(18)(19). Strains that could not be revitaliz...

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“…These probes were used to develop a DNA probe-based typing approach. The strain-specific DNA probes provide a simple binary output and have been presented before (13)(14)(15). We describe here the development of a new format for the binary typing technique.…”

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“…A serial dilution of labeled DNA from all strains (1 l) was spotted onto a membrane to check the labeling efficiency. The procedures for the generation, validation, and application of the 12 strain-specific DNA probes have been described in detail elsewhere (13)(14)(15). Purified DNA probes (n ϭ 12) were spotted (1 l) onto a membrane strip (5 by 80 mm; Hybond N ϩ ; Amersham Life Science, Little Chalfont, United Kingdom).…”

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Binary Typing of Staphylococcus aureus Strains through Reversed Hybridization Using Digoxigenin-Universal Linkage System-Labeled Bacterial Genomic DNA

Leeuwen

Libregts²,

Schalk³

et al. 2001

J Clin Microbiol

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A novel binary typing (BT) procedure, based on reversed hybridization of digoxigenin-universal linkage system-labeled bacterial DNA to strip-immobilized probes, is presented. Chromogenic detection of hybrids was performed. Staphylococcus aureus isolates (n ‫؍‬ 20) were analyzed to establish the feasibility of BT. A technically simple and fast procedure has been developed for application in routine microbiology laboratories.Reliable probe-based microbial typing systems are not yet commonplace in microbiological practice (1,4,7,8,12). In the past we identified domains that are differentially present within the staphylococcal genome on the basis of randomly amplified polymorphic DNA analysis. These probes were used to develop a DNA probe-based typing approach. The strain-specific DNA probes provide a simple binary output and have been presented before (13-15). We describe here the development of a new format for the binary typing technique. In the newly described procedure DNA is extracted from overnight Staph-

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Intercenter reproducibility of binary typing for Staphylococcus aureus

Cited by 20 publications

References 22 publications

DNA Macroarray for Identification and Typing of Staphylococcus aureus Isolates

DNA Macroarray for Identification and Typing of Staphylococcus aureus Isolates

Recognition of SCC mec Types According to Typing Pattern Determined by Multienzyme Multiplex PCR-Amplified Fragment Length Polymorphism Analysis of Methicillin-Resistant Staphylococcus aureus

Binary Typing of Staphylococcus aureus Strains through Reversed Hybridization Using Digoxigenin-Universal Linkage System-Labeled Bacterial Genomic DNA

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