Development of a Canadian standardized protocol for subtyping methicillin-resistant Staphylococcus aureus using pulsed-field gel electrophoresis

A panel of 24 methicillin-resistant Staphylococcus aureus strains was distr ibuted to 15 laboratories in Canada to evaluate their in-house pulsed-field gel electrophoresis (PFGE) protocols and interpretation criteria. Attempts to compare fingerprint images using computer-aided analysis were not succe ssful due to variability in individual laboratory PFGE protocols. In additi on, individual site interpretation of the fingerprint patterns was inadequa te, as 7 of 13 sites (54%) made at least one error in interpreting the fing erprints from the panel. A 2-day standardized PFGE protocol (culture to gel image) was developed and distributed to all of the sites. Each site was re quested to use the standardized protocol on five strains from the original panel. Thirteen sites submitted gel images for comparisons. The protocol de monstrated excellent reproducibility and allowed interlaboratory comparison s with Molecular Analyst DST software (Bio-Rad) and 1.5% band tolerance.