Article Figures & Data
Figures
- FIG. 1.
Comparison of the amino acid sequences of the VEB-1, VEB-1a, VEB-1b, VEB-2, and VEB-3 beta-lactamases from P. aeruginosa JES from Thailand, P. aeruginosa KU-1 and KU-2 from Kuwait, P. aeruginosa 16 from Thailand, and E. cloacae 39 from China. Dashes indicate identical amino acids. The arrow indicates the position of the putative cleavage site of the leader peptide. Numbering is according to Ambler.
- FIG. 2.
Schematic representation of the three integrons found in E. cloacae 39, 42, and 57 and the structure of the blaVEB-3 gene cassette. Gene cassettes are shown as boxes, with arrows indicating the orientation of transcription and black circles indicating the 59-base element. The 5′ conserved segment contains the intI1 gene, which encodes integrase. The ereA2 gene was interrupted by IS1247, which are indicated by filled and empty triangles, respectively.
- FIG. 3.
(a) Plasmid DNA profiles of clinical isolates E. cloacae 39 and 57; (b) Southern blots hybridized with an intI1-specific probe; (c) Southern blots hybridized with a blaVEB-1-like specific probe. Lanes A, V517 plasmid size marker; lanes M, 1-kb DNA marker; lanes 1, plasmid DNA profiles of E. cloacae 39; lanes 2, plasmid DNA profiles of E. cloacae 57; lanes 3, chromosomal DNA of E. cloacae 39; lanes 4, chromosomal DNA of E. cloacae 57.
- FIG. 4.
(a) rep-PCR profiles of E. cloacae clinical isolates with primer ERIC2; (b) rep-PCR profiles of E. cloacae clinical isolates with primer REP. Lanes M, DNA molecular size markers; lanes A to G, rep-PCR profiles of seven distinct types of E. cloacae clinical isolates, respectively. Subscript numerals: 1, ERIC2 type; 2, REP PCR type.
Tables
- TABLE 1.
Sequences of primers used for detection of bla genes or genotyping of strains
ESBL or region Primer Sequence Position GenBank accession no. or reference TEM TEM-A 5′-ATA AAA TTC TTG AAG AC-3′ 1-17 X54604.1 TEM-B 5′-TTA CCA ATG CTT AAT CA-3′ 1075-1059 SHV SHV-FOR 5′-TGGTTATGCGTTATATTCGCC-3′ 69-89 X98100.1 SHV-REV 5′-GCTTAGCGTTGCCAGTGCT-3′ 936-918 PRE-1 PER-1-FOR 5′-AATTTGGGCTTAGGGCAGAA-3′ 278-306 Z21957 PER-1-REV 5′-ATGAATGTCATTATAAAAGC-3′ 1211-1192 VEB-1 VEB-1-FOR 5′-CGACTTCCATTTCCCGATGC-3′ 226-245 AF220758 VEB1-REV 5′-GGACTCTGCAACAAATACGC-3′ 868-849 CTX-M-3 M13U 5′-GGTTAAAAAATCACTGCGTC-3′ 65-84 X92506 M13L 5′-TTGGTGACGATTTTAGCCGC-3′ 928-909 CTX-M-9 M9U 5′-ATGGTGACAAAGAGAGTGCA-3′ 1-20 AF252621.2 M9L 5′-CCC TTC GGC GAT GAT TCT C-3′ 870-852 CTX-M-2 M25U 5′-ATGATGACTCAGAGCATTCG-3′ 304-323 AJ416343.1 M25L 5′-TGGGTTACGATTTTCGCCGC-3′ 1169-1150 VEBcas VEBcas-F 5′-GTTAGCGGTAATTTAACCAGATAG-3′ 11-34 AY027870.1 VEBcas-B 5′-CGGTTTGGGCTATGGGCAG-3′ 1081-1063 Rep-PCR ERIC2 5′-AAGTAAGTGACTGGGGTGAGCG-3′ 22 REP 2-1 5′-ICG ICT TAT CIG GCC TAC-3′ 2 VEBINV1 5′-CAGTTTGAGCATTTGAATACAC-3′ 16 VEBINV2 5′-AGC GTA TTT GTT GCA GAG TCC-3′ Variable region of integron 5-CS 5′-GGC ATC CAA GCA GCA AGC-3′ 1163-1180 AF205943 3-CS 5′-AAG CAG ACT TGA CCT GAT-3′ 9231-9214 VEB clone VEBc-R 5′-CCGGGATCCATGAAAATCGTAAAAAGGATATTATT-3′ AF220758 VEBc-L 5′-CCCAAGCTTTTATTTATTCAAATAGTAATTCCACG-3 - TABLE 2.
MICs of selected antibiotics for representative E. cloacae clinical isolates
E. cloacae isolate Etest MIC (μg/ml)a IPM FEP AK CAZ CTX PTc CIP TZ-TZL FOX CT-CTL 72 0.38 3 12 48 32 4 0.13 >32/>4 >256 >16/>1 64 0.25 4 8 48 >256 2 0.25 >32/>4 >256 >16/>1 5 0.25 >256 2 >256 >256 >256 >32 >32/>4 >256 >16/>1 57 0.25 24 >256 >256 >256 >256 >32 >32/>4 >256 >16/>1 136 0.25 3 1.5 12 >256 6 0.75 >32/>4 >256 >16/>1 42 0.5 128 >256 >256 >256 >256 >32 >32/>4 >256 >16/>1 150 0.38 >256 >256 >256 >256 >256 >32 >32/>4 48 >16/>1 108 0.25 >256 >256 >256 >256 >256 >32 >32/>4 >256 >16/>1 39 0.75 >256 >256 >256 >256 >256 >32 >32/>4 >256 >16/>1 103 0.25 0.75 4 96 32 96 0.16 >32/>4 >256 >16/>1 212 0.5 2 >256 3 6 48 >32 >32/>4 >256 >16/>1 142 0.25 3 2 >256 >256 >256 0.06 >32/>4 >256 >16/>1 ↵ a Abbreviations: IPM, imipenem; FEP, cefepime; AK, amikacin; CAZ, ceftazidime; CTX, cefotaxime; PTc, cefoperazone-sulbactam; CIP, ciprofloxacin; TZ-TZL, ceftazidime-ceftazidime-clavulanic acid; FOX, cefoxitin; CT-CTL, cefotaxime-cefotaxime-clavulanic acid.
- TABLE 3.
Genotyping of rep-PCR products and beta-lactamase genes of 27 ceftazidime-resistant or cefotaxime-resistant E. cloacae clinical isolates and correlation with phenotypic ESBL tests
Type Isolate(s) Result for bla gene for: Result by NCCLS K-Ba method TEM-1 CTX-M-3 CTX-M-9a VEB-1 SHV-5a A 5, 25 − − + − − +b B 64 − − + − − + 12, 150 + − + − − − 33 + + − − + + 39, 40, 42, 71 + + − + − − 236, 57, 108 + + − + − − 77, 88 + + − + − − 157, 205, 189 + − − + − − 17, 224 + + − − − − C 72 + + − − − + D 103 − − − − − − E 136 + − + − − − 216 − − + + − − F 140, 212 − − − − − − G 142 + − − − − − - TABLE 4.
MICs of beta-lactams for recombinant bacteria E. coli EV3 and E. coli ET
E. coli strain Etest MIC (μg/ml)a IPM FEP CAZ CTX PTc CAZ-TZL FOX CTX-CTL AM EV3 0.16 3 >256 24 0.38 >32/0.25 1.5 8/0.25 >256 ET 0.125 0.032 0.47 0.16 0.38 0.5/0.094 1.5 0.5/0.094 2 ↵ a Abbreviations: IPM, imipenem; FEP, cefepime; CAZ, ceftazidime; CTX, cefotaxime; PTc, cefoperazone-sulbactam; TZL, ceftazidime-clavulanic acid; FOX, cefoxitin; CTL, cefotaxime-clavulanic acid, AM, ampicillin.
