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Background: Staphylococcus aureus is a common cause of infection, particularly in persons colonized by this organism. Virulent strains of methicillin-resistant S. aureus (MRSA) have emerged in the general community., Methods: A nationally representative survey of nasal colonization with S. aureus was conducted from 2001 through 2004 as part of the National Health and Nutrition Examination Survey. MRSA isolates were identified by the oxacillin disk-diffusion method. The pulsed-field gel electrophoresis (PFGE) type was determined for all MRSA isolates. A t statistic was used to compare the prevalence of colonization across biennia and across population subgroups. Cofactors independently associated with colonization were determined with backward stepwise logistic modeling., Results: The prevalence of colonization with S. aureus decreased from 32.4% in 2001-2002 to 28.6% in 2003-2004 (P < .01), whereas the prevalence of colonization with MRSA increased from 0.8% to 1.5% (P < .05). Colonization with MRSA was independently associated with healthcare exposure in males and with having been born in the United States, age > or =60 years, diabetes, and poverty in females. In 2003-2004, a total of 19.7% (95% confidence interval, 12.4%-28.8%) of MRSA-colonized persons carried a PFGE type associated with community transmission., Conclusions: Nasal colonization with MRSA has increased in the United States, despite an overall decrease in nasal colonization with S. aureus. PFGE types associated with community transmission only partially account for the increase in MRSA colonization.

Objectives: We conducted a retrospective study to determine trends and characteristics of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) in Hawaii., Methods: We reviewed medical records of patients with MRSA infections during July 2001-June 2003 in four healthcare facilities. A case was defined as a patient with MRSA infection (colonization excluded), diagnosed in ambulatory settings or < or = 48 h after hospitalization, without previous MRSA or healthcare risk factors. Pulsed-field gel electrophoresis (PFGE) and typing of resistance and toxin genes was performed in 40 MRSA isolates., Results: CA-MRSA infections increased from 28 (23% of MRSA infections) to 65 (32%) per quarter over the 2-year period (P<0.05). Pacific islanders accounted for 51% of 389 case-patients, but only 24% of the Hawaii population. In the pediatric hospital, Pacific Islanders represented 76% of 90 case-patients versus 35% of the hospital population. Hospital admission, required for 40% (154/389), was associated with prior antimicrobial treatment (P<0.01). The staphylococcal cassette chromosome mec type IV was detected in 38/40 isolates; 31 isolates carried Panton-Valentine leukocidin genes and 22 belonged to the same staphylococcal lineage., Conclusions: In Hawaii, prevention strategies for CA-MRSA infections should focus on Pacific Islanders. CA-MRSA infections in Hawaii appear to be related to strains causing disease throughout the United States.

Background: Staphylococcus aureus is a common cause of disease, particularly in colonized persons. Although methicillin-resistant S. aureus (MRSA) infection has become increasingly reported, population-based S. aureus and MRSA colonization estimates are lacking., Methods: Nasal samples for S. aureus culture and sociodemographic data were obtained from 9622 persons > or = 1 year old as part of the National Health and Nutrition Examination Survey, 2001-2002. After screening for oxacillin susceptibility, MRSA and selected methicillin-susceptible S. aureus isolates were tested for antimicrobial susceptibility, pulsed-field gel electrophoresis clonal type, toxin genes (e.g., for Panton-Valentine leukocidin [PVL]), and staphylococcal cassette chromosome mec (SCCmec) type I-IV genes., Results: For 2001-2002, national S. aureus and MRSA colonization prevalence estimates were 32.4% (95% confidence interval [CI], 30.7%-34.1%) and 0.8% (95% CI, 0.4%-1.4%), respectively, and population estimates were 89.4 million persons (95% CI, 84.8-94.1 million persons) and 2.3 million persons (95% CI, 1.2-3.8 million persons), respectively. S. aureus colonization prevalence was highest in participants 6-11 years old. MRSA colonization was associated with age > or = 60 years and being female but not with recent health-care exposure. In unweighted analyses, the SCCmec type IV gene was more frequent in isolates from participants of younger age and of non-Hispanic black race/ethnicity; the PVL gene was present in 9 (2.4%) of 372 of isolates tested., Conclusions: Many persons in the United States are colonized with S. aureus; prevalence rates differ demographically. MRSA colonization prevalence, although low nationally in 2001-2002, may vary with demographic and organism characteristics.

A highly stable strain of Staphylococcus aureus with a pulsed-field gel electrophoresis type of USA300 and multilocus sequence type 8 has been isolated from patients residing in diverse geographic regions of the United States. This strain, designated USA300-0114, is a major cause of skin and soft tissue infections among persons in community settings, including day care centers and correctional facilities, and among sports teams, Native Americans, men who have sex with men, and military recruits. The organism is typically resistant to penicillin, oxacillin, and erythromycin (the latter mediated by msrA) and carries SCCmec type IVa. This strain is variably resistant to tetracycline [mediated by tet(K)]; several recent isolates have decreased susceptibility to fluoroquinolones. S. aureus USA300-0114 harbors the genes encoding the Panton-Valentine leucocidin toxin. DNA sequence analysis of the direct repeat units within the mec determinant of 30 USA300-0114 isolates revealed differences in only a single isolate. Plasmid analysis identified a common 30-kb plasmid that hybridized with blaZ and msrA probes and a 3.1-kb cryptic plasmid. A 4.3-kb plasmid encoding tet(K) and a 2.6-kb plasmid encoding ermC were observed in a few isolates. DNA microarray analysis was used to determine the genetic loci for a series of virulence factors and genes associated with antimicrobial resistance. Comparative genomics between USA300-0114 and three other S. aureus lineages (USA100, USA400, and USA500) defined a set of USA300-0114-specific genes, which may facilitate the strain's pathogenesis within diverse environments.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) infection typically occurs in chronically ill patients requiring long-term antimicrobial therapy or hospitalization. However, community-associated MRSA (CA-MRSA) necrotizing soft tissue infections seem to be increasing in incidence. Our aim was to describe the incidence and microbiologic characteristics of CA-MRSA isolates collected at an army community hospital., Methods: We report a retrospective review of MRSA isolates identified during 1998-2003 at the microbiology laboratory of Moncrief Army Community Hospital that serves a community of approximately 40,000 transient residents yearly in Fort Jackson, South Carolina. We evaluated the incidence of MRSA in our laboratory during 1998-2003. For MRSA isolates from 2003, we evaluated antimicrobial susceptibility patterns. Six selected isolates were evaluated by molecular typing, resistance gene analysis, and toxin analysis., Results: During 1998-2003, 241 (23%) of 1041 S. aureus isolates identified at the hospital microbiology laboratory were resistant to methicillin. Of these 241 MRSA isolates, 223 were cultured from outpatients. The incidence of MRSA in our population increased from 12% of S. aureus isolates in 1998 to 43% in 2003. In 2003, MRSA was cultured from 76 different patients. Isolates of MRSA were often resistant to erythromycin (91%), although resistance to other agents was less common: Ciprofloxacin (14%), levofloxacin (14%), clindamycin (3%), tetracycline (3%), and trimethoprim sulfamethoxazole (1%). No isolates were resistant to vancomycin, gentamicin, nitrofurantoin, or rifampin. Six CA-MRSA isolates were compared by pulsed-field gel electrophoresis (PFGE). Five were PFGE type USA300, and one was PFGE type USA100, based on the U.S. Centers for Disease Control and Prevention (CDC) classification scheme. The five USA300 isolates carried SCCmec type IV, and the USA100 carried SCCmec II. None of the isolates were positive by PCR for genes encoding enterotoxins A-E and H, or toxic shock syndrome toxin (TSST-1), but the five USA300 isolates carried the gene coding for Panton-Valentine leukocidin toxin., Conclusions: The incidence of MRSA at our institution is increasing. Isolates of MRSA show resistance patterns and microbiologic characteristics consistent with CA-MRSA isolates from the United States. Clinicians should consider the possibility of CA-MRSA in patients with soft-tissue infections who do not respond to initial therapy with beta-lactam antimicrobial agents.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is an emerging cause of infections outside of health care settings. We investigated an outbreak of abscesses due to MRSA among members of a professional football team and examined the transmission and microbiologic characteristics of the outbreak strain., Methods: We conducted a retrospective cohort study and nasal-swab survey of 84 St. Louis Rams football players and staff members. S. aureus recovered from wound, nasal, and environmental cultures was analyzed by means of pulsed-field gel electrophoresis (PFGE) and typing for resistance and toxin genes. MRSA from the team was compared with other community isolates and hospital isolates., Results: During the 2003 football season, eight MRSA infections occurred among 5 of the 58 Rams players (9 percent); all of the infections developed at turf-abrasion sites. MRSA infection was significantly associated with the lineman or linebacker position and a higher body-mass index. No MRSA was found in nasal or environmental samples; however, methicillin-susceptible S. aureus was recovered from whirlpools and taping gel and from 35 of the 84 nasal swabs from players and staff members (42 percent). MRSA from a competing football team and from other community clusters and sporadic cases had PFGE patterns that were indistinguishable from those of the Rams' MRSA; all carried the gene for Panton-Valentine leukocidin and the gene complex for staphylococcal-cassette-chromosome mec type IVa resistance (clone USA300-0114)., Conclusions: We describe a highly conserved, community-associated MRSA clone that caused abscesses among professional football players and that was indistinguishable from isolates from various other regions of the United States., (Copyright 2005 Massachusetts Medical Society.)

In an effort to find a rapid, efficient, and reliable method of screening large numbers of bacterial isolates for specific antimicrobial resistance genes, we compared conventional PCR results to the results generated using the TaqMan 5' nuclease PCR kit in conjunction with an ABI Prism 7700 Sequence Detector for detecting the mecA gene in various species of staphylococci. DNA was extracted using two techniques. The first used a high-salt extraction method suitable for conventional PCR but resulted in a 7.2% rate of PCR inhibition with the TaqMan technique. PCR inhibition could be overcome by diluting samples 1:5 prior to testing. The second method used the Qiagen QIAamp Tissue Kit; no instances of PCR inhibition were encountered with this method. A total of 197 (96%) of the 206 samples with no inhibition showed agreement between the two methods. Eight of the nine disagreements were likely the result of low-level DNA cross contamination caused by frequent specimen handling. Target DNA in all eight of these samples was first detected in the initial tests only after >30 PCR cycles, and all were negative upon repeat testing even after 40 PCR cycles using freshly extracted DNA. Among those positive samples in agreement, target DNA was invariably detected before 30 PCR cycles. The TaqMan assay eliminated the need to load, run, stain, and read agarose gels and provided the advantage of instant detection of PCR product by laser-activated fluorescence. Thus, final results were obtained 2 h after PCR was initiated, as opposed to a requirement of 2 days to examine 96 samples by agarose gel electrophoresis.