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...g/ml) was added to the lysis solution for the preparation of S. aureus chromosomal DNA. Isolates with similar banding patterns and no more than three band differences were considered clonally related =-=(32)-=-. Isolates with banding patterns similar to clonally related strains but with no more than four band differences were considered subtypes of the clonal group. Once isolates were recognized as having i...

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...d from all three clinical centers. To ensure that the general clonality observed among the S. aureus endophthalmitis isolates was not attributable to a methicillin-resistant S. aureus (MRSA) genotype =-=(21)-=-, strains comprising each of the five S. aureus clonotypes were analyzed for the presence of the mecA antibiotic resistance determinant (8). Briefly, bacteria from a 0.5-ml suspension of bacterial cel...

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... enrichment for the cytolytic phenotype among endophthalmitis isolates compared with its occurrence among isolates from the gastrointestinal tracts of healthy subjects (0 to 17%; P , 0.028 [x2 test]) =-=(12, 15)-=-. All isolates comprising both EF1 and EF2 clonotypes were cytolytic. Relationship between clonality of endophthalmitis isolates and final visual outcome. In the present study, a number of S. aureus c...

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...ory strains in animal models of disease. These studies showed that cytolytic E. faecalis not only causes more fulminant disease but also renders the infection unresponsive to therapeutic intervention =-=(17, 18)-=-. The production of most secreted and cell surface proteins in S. aureus is coordinately controlled by chromosomal regulatory loci termed accessory gene regulator (agr) and staphylococcal accessory re...

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...for 2 days at 37°C. E. faecalis FA2-2 (pAM714) and plasmid-free E. faecalis FA2-2 were used as positive and negative controls, respectively, for the detection of both cytolytic phenotype and genotype =-=(14)-=-. Of the 28 E. faecalis endophthalmitis isolates collected for this study, 13 (46.4%) possessed the cylA gene, and all cylA-positive strains were phenotypically positive for cytolysin expression as in...

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...utable to a methicillin-resistant S. aureus (MRSA) genotype (21), strains comprising each of the five S. aureus clonotypes were analyzed for the presence of the mecA antibiotic resistance determinant =-=(8)-=-. Briefly, bacteria from a 0.5-ml suspension of bacterial cells in phosphate-buffered saline were lysed by boiling in a sealed tube for 10 min, followed by centrifugation (10,000 3 g for 1 min) to rem...

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... regulator (sar) (6, 16, 20). Mutant strains of S. aureus with insertional mutations in the sar and agr loci are attenuated in virulence in experimental endophthalmitis compared with parental strains =-=(3, 4)-=-. Since sar and agr affect the expression of 12 or more unrelated genes (6, 20), the staphylococcal toxin(s) that contributes most significantly to the severity of disease has not yet been identified....

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...cts of healthy volunteers, as previously reported (12, 15). This observation is consistent with an enrichment in the cytolysin genotype among E. faecalis clinical isolates from other anatomical sites =-=(12, 13, 15)-=-. Cytolysin is a toxin capable of lysing both eukaryotic and prokaryotic cells and is most commonly encoded by highly transmissible pheromone-responsive plasmids (15). Cytolysin has been shown previou...

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...lis cytolysin operon. The following oligonucleotide primers were selected from published sequences: 59 AAT GGA TAA TAT TTC AGA ATT TGA AGT 39 (cylA1) and 59 TTC CCA CGA AAA TTT TAT AAA CCC 39 (cylA2) =-=(9)-=-. Briefly, a suspension of each isolate was prepared by removing bacterial colonies from an overnight plate culture with a moistened sterile swab and resuspending them in 1 ml of sterile 10 mM Tris, p...

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...LE 2. Correlation between severity of outcome and S. aureus endophthalmitis clonotype Clonotype (n) Severity of outcome (no. [%] of patients) Pb 20/200 or worsea Better than 20/200a SA1 (10) 7 (70) 3 =-=(30)-=- 0.054 SA2 (4) 2 (50) 2 (50) ND SA3 (2) 1 (50) 1 (50) ND SA4 (3) 2 (66) 1 (33) ND SA5 (4) 3 (75) 1 (25) ND Total (clonal) (23) 15 (65) 8 (35) 0.036Other isolates (nonclonal) (16) 5 (31) 11 (69) a Fina...

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...skin, eyelid margins, or nares (22, 31). With PFGE used to identify clonal relationships between strains, it was recently shown that nasal colonization patterns by S. aureus and S. epidermidis differ =-=(10, 11)-=-. In the case of S. aureus, the same strain was observed to persistently colonize the host for periods of at least 2 years, while predominant S. epidermidis strains colonizing the nares were observed ...

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...ory strains in animal models of disease. These studies showed that cytolytic E. faecalis not only causes more fulminant disease but also renders the infection unresponsive to therapeutic intervention =-=(17, 18)-=-. The production of most secreted and cell surface proteins in S. aureus is coordinately controlled by chromosomal regulatory loci termed accessory gene regulator (agr) and staphylococcal accessory re...

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...used by more virulent organisms such as Staphylococcus aureus, Enterococcus faecalis, or Bacillus cereus, visual outcomes ranging from 20/ 100 to enucleation occur in approximately 50 to 90% of cases =-=(1, 5, 7)-=-. Despite a general association between visual outcome and the infectious agent, relatively little is known about the species-specific factors that account for the characteristic severity of each dise...

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... enrichment for the cytolytic phenotype among endophthalmitis isolates compared with its occurrence among isolates from the gastrointestinal tracts of healthy subjects (0 to 17%; P , 0.028 [x2 test]) =-=(12, 15)-=-. All isolates comprising both EF1 and EF2 clonotypes were cytolytic. Relationship between clonality of endophthalmitis isolates and final visual outcome. In the present study, a number of S. aureus c...

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...on of most secreted and cell surface proteins in S. aureus is coordinately controlled by chromosomal regulatory loci termed accessory gene regulator (agr) and staphylococcal accessory regulator (sar) =-=(6, 16, 20)-=-. Mutant strains of S. aureus with insertional mutations in the sar and agr loci are attenuated in virulence in experimental endophthalmitis compared with parental strains (3, 4). Since sar and agr af...

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...used by more virulent organisms such as Staphylococcus aureus, Enterococcus faecalis, or Bacillus cereus, visual outcomes ranging from 20/ 100 to enucleation occur in approximately 50 to 90% of cases =-=(1, 5, 7)-=-. Despite a general association between visual outcome and the infectious agent, relatively little is known about the species-specific factors that account for the characteristic severity of each dise...

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...2) 14 (48) TABLE 2. Correlation between severity of outcome and S. aureus endophthalmitis clonotype Clonotype (n) Severity of outcome (no. [%] of patients) Pb 20/200 or worsea Better than 20/200a SA1 =-=(10)-=- 7 (70) 3 (30) 0.054 SA2 (4) 2 (50) 2 (50) ND SA3 (2) 1 (50) 1 (50) ND SA4 (3) 2 (66) 1 (33) ND SA5 (4) 3 (75) 1 (25) ND Total (clonal) (23) 15 (65) 8 (35) 0.036Other isolates (nonclonal) (16) 5 (31) ...

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...erity of outcome (no. [%] of patients) Pb 20/200 or worsea Better than 20/200a SA1 (10) 7 (70) 3 (30) 0.054 SA2 (4) 2 (50) 2 (50) ND SA3 (2) 1 (50) 1 (50) ND SA4 (3) 2 (66) 1 (33) ND SA5 (4) 3 (75) 1 =-=(25)-=- ND Total (clonal) (23) 15 (65) 8 (35) 0.036Other isolates (nonclonal) (16) 5 (31) 11 (69) a Final best-corrected visual acuity. b By Pearson’s chi square test. ND, not done (the numbers of isolates i...

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...tcome and S. aureus endophthalmitis clonotype Clonotype (n) Severity of outcome (no. [%] of patients) Pb 20/200 or worsea Better than 20/200a SA1 (10) 7 (70) 3 (30) 0.054 SA2 (4) 2 (50) 2 (50) ND SA3 =-=(2)-=- 1 (50) 1 (50) ND SA4 (3) 2 (66) 1 (33) ND SA5 (4) 3 (75) 1 (25) ND Total (clonal) (23) 15 (65) 8 (35) 0.036Other isolates (nonclonal) (16) 5 (31) 11 (69) a Final best-corrected visual acuity. b By Pe...

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...used by more virulent organisms such as Staphylococcus aureus, Enterococcus faecalis, or Bacillus cereus, visual outcomes ranging from 20/ 100 to enucleation occur in approximately 50 to 90% of cases =-=(1, 5, 7)-=-. Despite a general association between visual outcome and the infectious agent, relatively little is known about the species-specific factors that account for the characteristic severity of each dise...

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...onotype Clonotype (n) Severity of outcome (no. [%] of patients) Pb 20/200 or worsea Better than 20/200a SA1 (10) 7 (70) 3 (30) 0.054 SA2 (4) 2 (50) 2 (50) ND SA3 (2) 1 (50) 1 (50) ND SA4 (3) 2 (66) 1 =-=(33)-=- ND SA5 (4) 3 (75) 1 (25) ND Total (clonal) (23) 15 (65) 8 (35) 0.036Other isolates (nonclonal) (16) 5 (31) 11 (69) a Final best-corrected visual acuity. b By Pearson’s chi square test. ND, not done (...

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...present study for S. aureus endophthalmitis isolates. In both S. aureus and S. epidermidis endophthalmitis, a likely source for the infecting organism is the periocular skin, eyelid margins, or nares =-=(22, 31)-=-. With PFGE used to identify clonal relationships between strains, it was recently shown that nasal colonization patterns by S. aureus and S. epidermidis differ (10, 11). In the case of S. aureus, the...