title: Comparison of automated repetitive-sequence-based polymerase chain reaction and spa typing versus pulsed-field gel electrophoresis for molecular typing of methicillin-resistant Staphylococcus aureus.
abstract: Automated repetitive polymerase chain reaction (PCR) (DiversiLab, bioMérieux, St. Laurent, Quebec, Canada) and single locus sequence typing of the Staphylococcus protein A (spa) gene with spa-type assignment by StaphType RIDOM software were compared to pulsed-field gel electrophoresis (PFGE) as the "gold standard" method for methicillin-resistant Staphylococcus aureus (MRSA) typing. Fifty-four MRSA isolates were typed by all methods: 10 of known PFGE CMRSA type and 44 clinical isolates. Correct assignment of CMRSA type or cluster occurred for 47 of 54 (87%) of the isolates when using a rep-PCR similarity index (SI) of ≥95%. Rep-PCR gave 7 discordant results [CMRSA1 (3), CMRSA2 (1), CMRSA4 (1), and CMRSA10 (2)], and some CMRSA clusters were not distinguished (CMRSA10/5/9, CMRSA 7/8, and CMRSA3/6). Several spa types occurred within a single PFGE or repetitive PCR types among the 19 different spa types found. spa type t037 was shared by CMRSA3 and CMRSA6 strains, and CMRSA9 and most CMRSA10 strains shared spa type t008. Time to results for PFGE, repetitive PCR, and spa typing was 3-4 days, 24 h, and 48 h, respectively. The annual costs of using spa or repetitive PCR were 2.4× and 1.9× higher, respectively, than PFGE but routine use of spa typing would lower annual labor costs by 0.10 full-time equivalents compared to PFGE. Repetitive PCR is a good method for rapid outbreak screening, but MRSA isolates that share the same repetitive PCR or PFGE patterns can be distinguished by spa typing.
authors.affiliation: Calgary Laboratory Services, Calgary, Alberta, Canada. deirdre.church@cls.ab.ca
authors.forename: Deirdre L
authors.initials: DL
authors.lastname: Church
authors.forename: Barbara L
authors.initials: BL
authors.lastname: Chow
authors.forename: Tracie
authors.initials: T
authors.lastname: Lloyd
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Gregson
mesh_terms: D016000:Cluster Analysis; D016521:Electrophoresis, Gel, Pulsed-Field / Q000191:economics / Q000379:methods*; D005838:Genotype; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000145:classification* / Q000235:genetics*; D017720:Molecular Epidemiology / Q000191:economics / Q000379:methods; D058889:Molecular Typing / Q000191:economics / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D013203:Staphylococcal Infections / Q000453:epidemiology / Q000382:microbiology; D013997:Time Factors