Article Figures & Data
Figures
- FIG. 1.
Schematic representation (not to scale) of Eap and the encoding gene, showing the amino acid (aa) composition consisting of a 30-aa signal peptide (S) and 110-aa residue domains (R1, R2, … RX) found in multiple copies. The oligonucleotide primer binding sites of EAP-CON1 (A1) and EAP-CON2 (A2) as well as of EAP-P3wt (B1) and EAP-P2wt (B2) are indicated in base pairs (bp); sizes correspond to EAP of the Newman D2C strain.
- FIG. 2.
Agarose gel (1%) electrophoresis patterns showing PCR products amplified with eap primers by use of genomic DNA of S. aureus ATCC 29213 (lanes 1 and 4), S. aureus Newman (lanes 2 and 5), and S. aureus Wood 46 (lanes 3 and 6) as well as genomic DNA of S. epidermidis DSM 20044 (lane 7), S. haemolyticus DSM 20263 (lane 8), S. hominis subsp. hominis DSM 20238 (lane 9), S. intermedius DSM 20373 (lane 10), and S. lugdunensis DSM 4804 (lane 11). Lane M, DNA molecular mass marker (1-kb/100-bp DNA ladder); lanes 1 to 3, eap gene amplified with primers EAP-P2wt and EAP-P3wt; lanes 4 to 11, eap gene amplified with newly designed primers EAP-CON1 and EAP-CON2 (see Table 2).
- FIG. 3.
Southern blot analysis of genomic DNA (A) and Northern blot of total RNA (B). Lanes 1 to 4, S. epidermidis; lane 5, S. carnosus subsp. carnosus; lane 6, S. haemolyticus; lane 7, S. hominis subsp. hominis; lanes 8 to 9, S. saprophyticus subsp. saprophyticus; lanes 10 to 12, S. aureus (lane 10, strain Newman; lane 11, strain CI7; lane 12, strain Wood 46). Blots were prepared as described in Materials and Methods.
- FIG. 4.
SDS-PAGE (A) and Western immunoblot (B) analysis of SDS surface protein extracts. Lane M, marker; lanes 1 to 3, S. aureus strains CI7 (lane 1), Wood 46 (lane 2), and Newman (lane 3); lanes 4 to 8, strains of non-S. aureus staphylococcal species S. epidermidis (lanes 4 and 5), S. carnosus subsp. carnosus (lane 6), S. haemolyticus (lane 7), and S. hominis subsp. hominis (lane 8). Blots were prepared as described in Materials and Methods.
Tables
- TABLE 1.
Non-S. aureus staphylococcal (sub-)species studied
Species and subspecies No. of clinical isolatesa Type and/or reference strain(s)b Total no. of isolatesc Blood Other S. arlettae 0 0 DSM 20672T 1 S. auricularis 0 0 DSM 20609T 1 S. capitis subsp. capitis 4 1 DSM 20326T 6 S. capitis subsp. urealyticus 0 0 DSM 6717T 1 S. caprae 0 5 DSM 20608T 6 S. carnosus subsp. carnosus 0 0 DSM 20501T 1 S. carnosus subsp. utilis 0 0 DSM 11676T 1 S. chromogenes 0 0 DSM 20454T 1 S. cohnii subsp. cohnii 4 2 DSM 20260T 7 S. cohnii subsp. urealyticus 0 0 DSM 6718T 1 S. condimenti 0 0 DSM 11674T 1 S. delphini 0 0 DSM 20771T 1 S. epidermidis 45 5 DSM 20044T 51 S. equorum subsp. equorum 0 0 DSM 20674T, DSM 20675 2 S. equorum subsp. linens 0 0 DSM 15097T 1 S. felis 0 0 DSM 7377T 1 S. fleurettii 0 0 DSM 13212T 1 S. gallinarum 0 0 DSM 20610T 1 S. haemolyticus 14 1 DSM 20263T 16 S. hominis subsp. hominis 9 0 DSM 20328T 10 S. hominis subsp. novobiosepticus 0 0 ATCC 700236T 1 S. hyicus 0 1 DSM 20459T 2 S. intermedius 1 10 DSM 20373T 12 S. kloosii 0 0 DSM 20676T 1 S. lentus 0 0 DSM 20352T 1 S. lugdunensis 1 19 DSM 4804T 21 S. lutrae 0 0 DSM 10244T 1 S. muscae 0 0 DSM 7068T 1 S. nepalensis 0 0 DSM 15150T, DSM 15151 2 S. pasteuri 0 0 DSM 10656T 1 S. pettenkoferi 4 1 CCUG 51270T 6 S. piscifermentans 0 0 DSM 7373T 1 S. pseudintermedius 0 0 LMG 22219T, LMG 22220, LMG 22221, LMG 22222 4 S. saccharolyticus 0 0 DSM 20359T 1 S. saprophyticus subsp. bovis 0 0 CCM 4410T 1 S. saprophyticus subsp. saprophyticus 2 12 DSM 20229T 15 S. schleiferi subsp. coagulans 0 0 DSM 6628T 1 S. schleiferi subsp. schleiferi 0 5 DSM 4807T 6 S. sciuri subsp. carnaticus 0 0 ATCC 700058T 1 S. sciuri subsp. rodentium 0 0 ATCC 700061T 1 S. sciuri subsp. sciuri 2 1 DSM 20345T 4 S. simulans 1 4 DSM 20322T 6 S. succinus subsp. casei 0 0 DSM 15096T 1 S. succinus subsp. succinus 0 0 DSM 14617T 1 S. vitulinus 0 0 ATCC 51145T 1 S. warneri 5 0 DSM 20316T 6 S. xylosus 1 4 DSM 20266T 6 Total 93 71 52 216 ↵ a Isolates from blood and from other specimens (skin, wound, respiratory tract, urogenital tract, and venous catheters).
↵ b Type or reference strains of the respective (sub-)species as deposited in the American Type Culture Collection (ATCC); Czech Collection of Microorganisms (CCM); Culture Collection, University of Göteborg, Sweden (CCUG); German Collection of Microorganisms and Cell Cultures (DSM); or Belgian Coordinated Collections of Microorganisms (LMG).
↵ c Values represent total numbers of clinical isolates plus type and/or reference strain isolates.
- TABLE 2.
eap-targeting oligonucleotide primers used in this study
Name Oligonucleotide sequence (5′→3′) Amplicon size (bp) Source or reference EAP-CON1 TAC TAA CGA AGC ATC TGC C 230 This study EAP-CON2 TTA AAT CGA TAT CAC TAA TAC CTC EAP-P3wt GCA GCT AAG CCA TTA GAT AAA TCA TCA AGT TGC TTA CAC C Variablea Derived from primer P3 (19)b EAP-P2wt TTA AAA TTT AAT TTC AAT GTC TAC TTT TTT AAT GTC
