Ribotyping of<i>Staphylococcus aureus</i>: an assessment using well–defined strains

Richardson, Judith; Aparicio, Pilar; Marples, R. R.; Cookson, Barry

doi:10.1017/s0950268800057459

Cited by 14 publications

(12 citation statements)

References 28 publications

(27 reference statements)

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“…Isolates of MRSA from France (QC01 and QC07) and Spain (211 and 212) have given similar patterns on ribotyping, PFGE (2,34), and coagulase typing (this study) and are thought to be representative of an epidemic clone circulating within Europe. In contrast, the German isolate 94/14013 was distinct and clearly separable from the other European isolates studied (2, 41) ( Fig.…”

Section: Discussionmentioning

confidence: 60%

“…The propagating strains, PS 42E and PS 71, represent some of the diverse types of isolates from human clinical material. They can be differentiated by ribotype (34) and by their unique coagulase RFLP patterns, 9 and 10, respectively (Fig. 2).…”

Section: Discussionmentioning

confidence: 99%

“…Bacterial strains were examined under a code that was broken upon completion of the analysis of the results. Eighty-five S. aureus strains representing EMRSA-1 to -16, including the original Jevons strain (NCTC 10442) and two duplicates, together with 10 methicillin-sensitive S. aureus propagating strains (PS), were studied (Table 1) (2,12,31,33,34,41 Bacteriophage typing. This was done by the method described by Blair and Williams (5) at the RTD and a 100ϫ RTD with the current set of international phages (3) and supplementary phages (32).…”

Section: Methodsmentioning

confidence: 99%

“…Recently, several investigators have described DNA-based techniques for typing strains (13,17,34,40). Pulsed-field gel electrophoresis (PFGE) is now recognized as being the most discriminatory method for gene typing strains of S. aureus, and it has been used to investigate nosocomial outbreaks (4,39).…”

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confidence: 99%

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Molecular Typing of Staphylococcus aureus Based on PCR Restriction Fragment Length Polymorphism and DNA Sequence Analysis of the Coagulase Gene

Richardson²,

1998

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A typing procedure for Staphylococcus aureus was developed based on improved PCR amplification of the coagulase gene and restriction fragment length polymorphism (RFLP) analysis of the product. All coagulasepositive staphylococci produced a single PCR amplification product of either 875, 660, 603, or 547 bp. Those strains of epidemic methicillin-resistant S. aureus 16 (EMRSA-16) studied all gave a product of 547 bp. PCR products were digested with AluI and CfoI, and the fragments were separated by gel electrophoresis. Ten distinct RFLP patterns were found among 85 isolates of methicillin-resistant S. aureus (MRSA) and 10 propagating strains (PS) of methicillin-sensitive S. aureus (MSSA) examined. RFLP patterns 1, 2, and 3 were specific to strains of EMRSA-3, -15, and -16, respectively. By contrast, RFLP patterns 4 and 5 were seen with a heterogeneous collection of strains, together with drug-resistant forms of S. aureus isolated in Europe and four propagating strains used for the international phage set. RFLP pattern 6 was given by the Airedale isolate and PS 95. RFLP pattern 7 encompassed EMRSA-2 (isolate 331), PS 94, and PS 96. An isolate from Germany gave RFLP pattern 8. Eight strains of MSSA gave patterns similar to those of methicillin-resistant strains (RFLP patterns 3, 4, 5, 6, and 7), but two, PS 42E and PS 71, gave unique RFLP patterns 9 and 10, respectively. The coagulase gene PCR products for 24 isolates of MRSA and two isolates of MSSA were sequenced for both strands. The sequences were aligned, and evolutionary lineages were inferred based on pairwise distances between isolates.

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Section: Discussionmentioning

confidence: 60%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Molecular Typing of Staphylococcus aureus Based on PCR Restriction Fragment Length Polymorphism and DNA Sequence Analysis of the Coagulase Gene

Richardson²,

1998

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“…Many molecular methods are available to type MRSA strains, including pulsed-field gel electrophoresis (PFGE) (7), PCR assays for rapidly evolving repeat elements (8), randomly amplified polymorphic DNA (RAPD) analysis (9), multilocus sequence typing (MLST) (10), typing of the polymorphic spa gene (11)(12)(13), ribotyping (14), mecA:Tn554 probe typing (15), staphylococcal cassette chromosome mec element (SCCmec) typing (16), multilocus variable-number tandem-repeat analysis (MLVA) (17)(18)(19), and DNA macroarray (20). These approaches have individual strengths and weaknesses.…”

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confidence: 99%

Whole-Genome Sequencing for High-Resolution Investigation of Methicillin-Resistant Staphylococcus aureus Epidemiology and Genome Plasticity

et al. 2014

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Methicillin-resistant Staphylococcus aureus (MRSA) infections pose a major challenge in health care, yet the limited heterogeneity within this group hinders molecular investigations of related outbreaks. Pulsed-field gel electrophoresis (PFGE) has been the gold standard approach but is impractical for many clinical laboratories and is often replaced with PCR-based methods. Regardless, both approaches can prove problematic for identifying subclonal outbreaks. Here, we explore the use of whole-genome sequencing for clinical laboratory investigations of MRSA molecular epidemiology. We examine the relationships of 44 MRSA isolates collected over a period of 3 years by using whole-genome sequencing and two PCR-based methods, multilocus variablenumber tandem-repeat analysis (MLVA) and spa typing. We find that MLVA offers higher resolution than spa typing, as it resolved 17 versus 12 discrete isolate groups, respectively. In contrast, whole-genome sequencing reproducibly cataloged genomic variants (131,424 different single nucleotide polymorphisms and indels across the strain collection) that uniquely identified each MRSA clone, recapitulating those groups but enabling higher-resolution phylogenetic inferences of the epidemiological relationships. Importantly, whole-genome sequencing detected a significant number of variants, thereby distinguishing between groups that were considered identical by both spa typing (minimum, 1,124 polymorphisms) and MLVA (minimum, 193 polymorphisms); this suggests that these more conventional approaches can lead to false-positive identification of outbreaks due to inappropriate grouping of genetically distinct strains. An analysis of the distribution of variants across the MRSA genome reveals 47 mutational hot spots (comprising ϳ2.5% of the genome) that account for 23.5% of the observed polymorphisms, and the use of this selected data set successfully recapitulates most epidemiological relationships in this pathogen group.

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