Comparison of four genotyping assays for epidemiological study of methicillin-resistantStaphylococcus aureus

Abstract

Twenty-six methicillin-resistant Staphylococcus aureus strains were genetically differentiated by interrepeat PCR and the results compared with those of ribotyping, pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA analysis obtained in a previous study for the same strains. The comparison showed that the PCR-mediated assays were as discriminatory as PFGE, whereas ribotyping was the least powerful genotyping method. Due to the ease of performance, PCR fingerprinting may become the method of choice for establishing clonal relationship among Staphylococcus aureus isolates.

Comparison of an improved RAPD fingerprinting with different typing methods for discriminating clinical isolates of Staphylococcus spp.

“…A variety of typing systems for differentiating among strains of staphylococci, particularly methicillin resistant Staphylococcus aureus (MRSA) and coagulasenegative staphylococci, can be valuable tools for assessing the clonal nature of bacteria involved in nosocomial outbreaks (see [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16] and their references). The more widely used methods based on phenotypic assays have limitations in discriminatory power [15].…”

“…Recently, techniques based on polymerase chain reaction (PCR) have provided an additional tool for a Staphylococcus spp. typing, starting from small quantities of DNA [9,10,14]. The characterization of mec gene [5] and of the 16S-23S rDNA spacer region [16] indicates that the PCR-amplification of these sequences could be useful for MRSA typing.…”

“…This technique was also successfully applied in order to subtype Staphylococcus spp. of clinical interest [6][7][8][9][10]. However the RAPD method has its drawbacks.…”

“…We have studied different parameters of the amplification reaction to improve the reproducibility and the discriminative value of the method. The improved RAPD fingerprinting was compared with biotyping, antibiotic assays, plasmid profiles, and REA patterns since relatively little efforts has been devoted to apply all these types of techniques to the same set of bacterial strains [10,15]. Such a study provides experimental data to assess the utility of these different methods for the characterization of clinical isolates of Staphylococcus spp.…”

Use of an Automated Multiple-Locus, Variable-Number Tandem Repeat-Based Method for Rapid and High-Throughput Genotyping of Staphylococcus aureus Isolates

“…Arbitrarily primed PCR methods are less expensive in terms of reagent costs (21), but they also suffer from poor interlaboratory reproducibility. Ribotyping is limited to use with bacterial species that contain several copies of the ribosomal locus (38) and has a lower interstrain discriminatory power than the other techniques (41). Multilocus sequence typing (MLST) shares the advantages of PCR-based methodologies, and the variability of the data obtained by MLST appears to be limited.…”