Molecular typing and clonal relatedness

LR Laura Ruiz-Ripa
CS Carmen Simón
SC Sara Ceballos
CO Carmelo Ortega
MZ Myriam Zarazaga
CT Carmen Torres
EG Elena Gómez-Sanz
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All SA were subjected to spa-typing by PCR and amplicon sequencing, and the obtained sequences were analyzed using Ridom Staph-Type software version 1.5.21 (Ridom GmbH, Münster, Germany) [32]. Multi Locus Sequence Typing (MLST) was performed in all SA isolates [32], and according to the sequence-type (ST), the isolates were ascribed to the different clonal complexes (CC). MLST of SP isolates was likewise performed as previously described [33]. All isolates were characterized by agr-typing following standard methodology [32, 33]. SCCmec-typing was undergone in MRSP and MRSA as previously described [13, 34].

PFGE of total DNA restricted with SmaI enzyme was performed on MRSP as previously described [35]. Isolates were considered different clones when they exhibited more than three bands of difference in PFGE band patterns and subclones when PFGE band patterns differed between 1 and 3 bands [36].

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