Mbo 1

Epidemiological types of all A. baumannii isolates were determined by Amplified Fragment Length Polymorphism (AFLP) (19 (link)). Briefly, genomic DNA from isolates were double digested with Mbo I and Mse I (Fermentas, Lithuania) and ligated to their corresponding adaptors. Ligation products served as templates for preliminary-PCR amplification using PreAmp-Mbo and PreAmp-Mse primers. Diluted preliminary-PCR products were then used for selective PCR amplification to generate AFLPs profiles. To determine the AFLP genotypes, the selective PCR products were separated by electrophoresis on 2% (w/v) agarose gels. Gel images were analyzed by BioNumerics version 5.10 (Applied Maths) using A. baumannii NCTC 12156 as a normalization reference. The similarity between the band patterns was calculated using the Dice coefficient (with an optimization of 0.5% and a position tolerance of 1%). The AFLP clusters and type identification were defined by groups formed at 60% and 90% Dice similarity cutoffs, respectively, on a dendrogram constructed by the unweighted-pair group method using average linkages (UPGMA) (19 (link)).