Repeat-based subtyping and grouping of Staphylococcus aureus from human infections and bovine mastitis using the R-domain of the clumping factor A gene

Abstract

Staphylococcus aureus has become an emerging public health concern. Markers capable of differentiating separate host-specific lineages are needed for tracing strain sources. Thus, a coding variable number tandem repeat-based typing was explored in this study, based on R-domain of clumping factor A (clfA) gene. DNA from isolates and strains of human infections and bovine mastitis were amplified and sequenced. Sequences of clfA from published strains were also analyzed. Results indicate that except one with 36 copies, 44 of the 55 R-domains had repeat copies between 44 and 57, whereas the remaining 10 had 59.5 to 73 copies. Furthermore, human isolates were polymorphic, while mastitis isolates were clonal. Phylogenetic grouping assigned host-specific strains into respective clusters. The repeats were stable during passages in milk, nutrient broth, and invasion of mammary cells showing suitability for typing. Our data show that the R-domain can be useful for typing and grouping host-specific lineages. Moreover, existence of variant repeats in human strains and the dominance of a clonal motif in mastitis may imply that a specific selection has occurred in the mammary gland.