RESUMO
BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) sequence type (ST) 45 was first reported in Taiwan in 2006. Since then, the prevalence of ST45 MRSA in clinical isolates has increased. This study was carried out to understand the changes in the proportions, evolutionary relationships, and infection advantages of ST45 and its related clones. MATERIALS AND METHODS: S. aureus including MRSA and MSSA (methicillin-sensitive S. aureus), and clonal complex (CC) 45 blood isolates were collected in 2000, 2005, and from January 2010 to August 2014. Molecular typing, multiple-locus variable-number tandem repeat analysis (MLVA) and single nucleotide polymorphism (SNP)-based phylogenetic analysis were performed. Fitness and virulence analyses were used to understand the infection advantages of the isolates. RESULTS: Among the 67 CC45 isolates, only MSSA ST508 isolates were found in 2000 and 2005. Since 2010, the prevalence of MRSA has increased, t1081/ST45 has become dominant, and MRSA ST508 has been found. Phylogenetic analysis indicated that most of the ST45 isolates were located in a cluster distinct from those of ST508 and ST929. However, the t026 isolates clustered with the ST508 isolates rather than with the other ST45 isolates. Moreover, fitness and virulence analyses revealed that the t1081 isolates had higher hemolytic activity than the t026 and ST508 isolates did. CONCLUSION: Our findings indicated that the increased prevalence of ST45 MRSA isolates from blood cultures in Taiwan was due to the t1081 isolates, and their high hemolytic activity may provide an infection advantage.
RESUMO
BACKGROUND: α-Hemolysin, encoded by hla, is a major virulence factor of Staphylococcus aureus. Sequence type (ST) 45 is a globally spread clone with increasing clinical prevalence in Taiwan. Our previous study showed that among the CC45 isolates, the spa type t1081 isolates presented greater hemolytic activity. MATERIALS AND METHODS: The hemolytic activity of 67 CC45 isolates (44 t1081 and 23 non-t1081) from clinical blood cultures was assessed using rabbit red blood cells. The sequences of hla and its upstream regulatory regions and RNAIII were compared between the two groups. The expression of hla and its regulators RNAIII, mgrA, and saeR was analyzed via qRTâPCR, while Hla protein levels were measured via Western blotting. RESULTS: Compared with non-t1081 isolates, t1081 isolates presented increased hemolytic activity. No significant differences in hla sequences, upstream regulatory regions, or gene expression levels were detected between the two groups. The expression of the transcriptional regulators mgrA and saeR was also similar between the two groups. Western blotting revealed increased Hla protein in the t1081 isolates. However, neither the sequence or expression of RNAIII, a regulator of hla at both the transcriptional and posttranscriptional levels, differed between the groups. CONCLUSION: Our study revealed that, compared with other CC45 isolates, the t1081/ST45 isolates presented greater hemolytic activity. This heightened activity was due mainly to increased Hla protein levels. Moreover, the higher translation levels may be independent of the known regulator RNAIII, indicating a potential RNAIII-independent mechanism for Hla regulation.