2000
DOI: 10.1089/mdr.2000.6.189
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Molecular Typing of Methicillin-ResistantStaphylococcus aureusby Pulsed-Field Gel Electrophoresis: Comparison of Results Obtained in a Multilaboratory Effort Using Identical Protocols and MRSA Strains

Abstract: Pulsed-field gel electrophoresis (PFGE) has become the gold standard of molecular methods in epidemiological investigations. In spite of its high resolving power, use of the method has been hampered by inadequate laboratory-to-laboratory reproducibility. In the project described here we have addressed this problem by organizing a multilaboratory effort in which the same bacterial strains (subtype variants of the Iberian and Brazilian methicillin-resistant Staphylococcus aureus--MRSA--clones) were analyzed by t… Show more

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Cited by 273 publications
(191 citation statements)
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“…1) were performed by the semi-automatic VITEK2 system (bioMérieux, SA, France) and the disk diffusion method for clindamycin [11]. All isolates were tested by polymerase chain reaction (PCR) for the presence of mecA [12] and were also characterized by pulsed-field gel electrophoresis (PFGE) [13]. The resulting SmaI patterns were analyzed by both visual inspection and automatically with BioNumerics software version 4.61 (Applied Maths, Sint-Martens-Latem, Belgium) [14,15].…”
mentioning
confidence: 99%
“…1) were performed by the semi-automatic VITEK2 system (bioMérieux, SA, France) and the disk diffusion method for clindamycin [11]. All isolates were tested by polymerase chain reaction (PCR) for the presence of mecA [12] and were also characterized by pulsed-field gel electrophoresis (PFGE) [13]. The resulting SmaI patterns were analyzed by both visual inspection and automatically with BioNumerics software version 4.61 (Applied Maths, Sint-Martens-Latem, Belgium) [14,15].…”
mentioning
confidence: 99%
“…In principle, the results generated should lead to good reproducibility and to comparability between centers, although this has yet to be formally demonstrated. This contrasts with PFGE, which has been shown to be poorly reproducible between centers (4,6,15,19), leading to considerable effort to introduce standardized protocols to improve reproducibility and technical quality (3,15). However, MLST is expensive and technically demanding, and it is important to establish whether it represents a clear advantage in terms of discrimination.…”
mentioning
confidence: 99%
“…However, the majority of these isolates showed no resistance to methicillin. The isolates which were resistant to methicillin had a further copy of mecA, identical to that of MRSA isolates (46,309).…”
Section: Evolution Of Mrsamentioning
confidence: 86%
“…In addition, international comparison of MRSA strains has become increasingly relevant because of the pandemic spread of MRSA strains. Despite established interpretation criteria (262), availability of sophisticated software packages, and several inter-laboratory standardization initiatives for PFGE (46,281), inter-laboratory reproducibility is difficult to achieve. Although software which transform bands mathematically to numerical values do exist, PFGE and other banding pattern based methods are prone to bias due to manual selection of bands.…”
Section: Molecular Typing Of Mrsamentioning
confidence: 99%