title: Molecular typing of methicillin-resistant Staphylococcus aureus by 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 twenty investigators in thirteen different laboratories according to an indentical protocol, which is reproduced here in detail. PFGE patterns obtained were analyzed at a central laboratory in order to identify specific technical problems that produced substandard macrorestriction patterns. The results including the specific technical problems and their most likely causes are described in this communication. Also listed are seven major epidemic clones of MRSA which have been characterized by molecular fingerprinting techniques and the prototypes of which have been deposited at the American Type Culture Collection, from where they will be available for interested investigators for the purpose of typing MRSA isolates. It is hoped that this communication will contribute to the improvement of the reproducibility and technical/aesthetic quality of PFGE analysis.
authors.affiliation: Laboratory of Microbiology, The Rockefeller University, New York, NY 10021, USA.
authors.forename: M
authors.initials: M
authors.lastname: Chung
authors.forename: H
authors.initials: H
authors.lastname: de Lencastre
authors.forename: P
authors.initials: P
authors.lastname: Matthews
authors.forename: A
authors.initials: A
authors.lastname: Tomasz
authors.forename: I
authors.initials: I
authors.lastname: Adamsson
authors.forename: M
authors.initials: M
authors.lastname: Aires de Sousa
authors.forename: T
authors.initials: T
authors.lastname: Camou
authors.forename: C
authors.initials: C
authors.lastname: Cocuzza
authors.forename: A
authors.initials: A
authors.lastname: Corso
authors.forename: I
authors.initials: I
authors.lastname: Couto
authors.forename: A
authors.initials: A
authors.lastname: Dominguez
authors.forename: M
authors.initials: M
authors.lastname: Gniadkowski
authors.forename: R
authors.initials: R
authors.lastname: Goering
authors.forename: A
authors.initials: A
authors.lastname: Gomes
authors.forename: K
authors.initials: K
authors.lastname: Kikuchi
authors.forename: A
authors.initials: A
authors.lastname: Marchese
authors.forename: R
authors.initials: R
authors.lastname: Mato
authors.forename: O
authors.initials: O
authors.lastname: Melter
authors.forename: D
authors.initials: D
authors.lastname: Oliveira
authors.forename: R
authors.initials: R
authors.lastname: Palacio
authors.forename: R
authors.initials: R
authors.lastname: Sá-Leão
authors.forename: I
authors.initials: I
authors.lastname: Santos Sanches
authors.forename: J H
authors.initials: JH
authors.lastname: Song
authors.forename: P T
authors.initials: PT
authors.lastname: Tassios
authors.forename: P
authors.initials: P
authors.lastname: Villari
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods / Q000592:standards; D006801:Humans; D007753:Laboratories / Q000592:standards*; D016106:Methicillin Resistance; D008829:Microbiology; D012015:Reference Standards; D015203:Reproducibility of Results; D013203:Staphylococcal Infections / Q000453:epidemiology / Q000382:microbiology; D013211:Staphylococcus aureus / Q000145:classification* / Q000187:drug effects / Q000235:genetics*