We report the complete nucleotide sequence and analysis of pETB TY825 , a Staphylococcus aureus TY825 plasmid encoding exfoliative toxin B (ETB). S. aureus TY825 is a clinical isolate obtained from an impetigo patient in 2002. The size of pETB TY825 , 60.6 kbp, was unexpectedly larger than that of the archetype pETB TY4 (ϳ30 kbp). Genomic comparison of the plasmids shows that pETB TY825 has the archetype pETB TY4 as the backbone and has a single large extra DNA region of 22.4 kbp. The extra DNA region contains genes for resistance to aminoglycoside [aac(6=)/aph(2؆)], macrolide (msrA), and penicillin (blaZ). A plasmid deletion experiment indicated that these three resistance elements were functionally active. We retrospectively examined the resistance profile of the clinical ETB-producing S. aureus strains isolated in 1977 to 2007 using a MIC determination with gentamicin (GM), arbekacin (ABK), and erythromycin (EM) and by PCR analyses for aac(6=)/aph(2؆) and msrA using purified plasmid preparations. The ETB-producing S. aureus strains began to display high resistance to GM, which was parallel with the detection of aac(6=)/aph(2؆) and mecA, after 1990. Conversely, there was no significant change in the ABK MIC during the testing period, although it had a tendency to slightly increase. After 2001, isolates resistant to EM significantly increased; however, msrA was hardly detected in ETB-producing S. aureus strains, and only five isolates were positive for both aac(6=)/aph(2؆) and msrA. In this study, we report the emergence of a fusion plasmid carrying the toxin gene etb and drug resistance genes. Prevalence of the pETB TY825 carrier may further increase the clinical threat, since ETB-producing S. aureus is closely related to more severe impetigo or staphylococcal scalded-skin syndrome (SSSS), which requires a general antimicrobial treatment.
Exfoliative toxin (ET) is an exotoxin produced by staphylococcal species, causing blisters on human and animal skin (1). ET-producing Staphylococcus aureus is involved in staphylococcal scalded-skin syndrome (SSSS) or Ritter disease and in bullous impetigo in neonates (1-3). Serologically, ETs causing diseases in human have been divided into three major serotypes: ETA, ETB, and ETD (4-6). All types cause intraepidermal cleavage in the granular layer, without epidermal necrolysis or inflammatory response in the skin (4, 5, 7). ETs are serine proteases that selectively cleave desmoglein 1, a desmosomal protein connecting epidermal cells present in the epidermis (8).Virulence factors of staphylococci such as ET are accessory proteins, which are not essential for cell growth or division. Genetic determinants for these factors are often associated with mobile genetic elements, such as phages, plasmids, and pathogenicity islands (9-11). The eta gene is located on the genome of a temperate phage ( ETA) (12), the etb gene is on a large plasmid (4, 13), and the etd gene is chromosomally located in a pathogenicity island (6).We previously reported the complete nucleotide sequence of th...