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. 2010 Feb 9:10:40.
doi: 10.1186/1471-2180-10-40.

PFGE diversity within the methicillin-resistant Staphylococcus aureus clonal lineage ST398

Thijs Bosch  1 , Albert J de Neeling, Leo M Schouls, Kim W van der Zwaluw, Jan A J W Kluytmans, Hajo Grundmann, Xander W Huijsdens
Affiliations

PFGE diversity within the methicillin-resistant Staphylococcus aureus clonal lineage ST398

Thijs Bosch et al. BMC Microbiol. .

Abstract

Background: Livestock has recently been identified as a new reservoir of methicillin-resistant Staphylococcus aureus (MRSA). Most isolates belong to ST398 and are non-typeable with PFGE using SmaI, making it difficult to study transmission and outbreaks. Therefore, a new PFGE using Cfr9I, a neoschizomer of SmaI was optimized and evaluated to investigate ST398 isolates.

Results: After optimizing and evaluating the Cfr9I PFGE, clear and reproducible banding patterns were obtained from all previously non-typeable MRSA (NT(SmaI) -MRSA) isolates. The PFGE patterns of ST398 isolates showed more diversity than with spa-typing and/or MLST. The PFGE results showed diversity within and between the two most prevalent spa-types of NT(SmaI) -MRSA (t011 and t108). No match was found, when comparing banding patterns of the NT(SmaI) -MRSA with 700 different PFGE types, obtained with SmaI digestion, in our database of more than 4000 strains. Furthermore, possible transmission among veterinarians and their family members was investigated and an outbreak of ST398 MRSA in a residential care facility was confirmed with the Cfr9I PFGE.

Conclusions: The adjusted PFGE can be used as a method for selecting important and distinct ST398 isolates for further research. The adjustments in the PFGE protocol using Cfr9I are easy to implement to study the ST398 clonal lineage in laboratories which already have a PFGE facility.

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Figures

Figure 1
Figure 1
Dendrogram of the Cfr9I PFGE results of NTSmaI-MRSA isolates with the 2 most prevalent spa-types in the Netherlands.
Figure 2
Figure 2
PFGE patterns of ST398 isolates digested with Cfr9I restriction enzyme using NCTC 8325 as the reference standard. Lanes 6, 12, 18, and 24, NCTC 8325; Lanes 1-5, isolates from an outbreak in a residential care facility, all PFGE pattern J; Lanes 7-8, and 14-15, two pairs of a veterinarian and a close family member with distinct PFGE patterns; Lanes 9-11, and 13, two pairs of a veterinarian and a close family member with identical banding patterns; Lanes 16-17, and 19-22, isolates of pig farm 6 with four different PFGE patterns; Lanes 23, and 25-28, isolates from pig farm 9 with identical banding patterns
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