Article Figures & Data
Figures
- Fig. 1.
Dendrogram of pulsed-field gel electrophoresis data (SpeI digests) showing genetic relatedness between the 10 KPC-producing reference isolates. M, molecular weight.
- Fig. 2.
(A) Plasmid extractions from cultures of the different isolates by the Kieser method. (B) Southern hybridization carried out with an internal probe for the blaKPC-2 gene. Lane 1, P. aeruginosa PA-1; lane 2, P. aeruginosa PA-2; lane 3, P. aeruginosa PA-3; lane 4, P. aeruginosa PA-4; lane 5, P. aeruginosa PA-5; lane 6, P. aeruginosa PA-6; lane 7, P. aeruginosa PA-7; lane 8, P. aeruginosa PA-8; lane 9, P. aeruginosa PA-9; lane 10, P. aeruginosa PA-10; lane 11, E. coli 50192 harboring four (7-, 48-, 66-, and 154-kb) plasmids. C, chromosome.
- Fig. 3.
Schematic representations of genetic structures surrounding the blaKPC gene in different P. aeruginosa isolates. (A) P. aeruginosa PR280 harboring blaKPC-5 from Puerto Rico (19); upstream of the blaKPC-5 gene, a portion of the transposable element Tn5563, previously described in plasmid pRA2 from P. alcaligenes, was found. (B) Novel structure characterized in PA-2 from Colombia. (C) Structure of Tn4401b. The dotted rectangles indicate the common regions between the different structures. Horizontal arrows indicate genes and their corresponding transcription orientations. Gray triangles represent the left inverted repeat (IRL) and right inverted repeat (IRR) of Tn4401. Small and empty triangles represent the inverted repeats of ISKpn6 and ISKpn7. Target site duplications (TSDs) are indicated above the sequence. Small black triangles above the sequence represent the primers listed in Table 2.
Tables
- Table 1.
Origin, structure of Tn4401, other β-lactamases, plasmid analysis, pulsotype, and sequence type of the P. aeruginosa isolates
Isolate Origin in Columbia Yr of isolation Site of isolation Location PCR and sequencing result (Tn4401 homology) blaKPC-2-positive plasmid Bacterial isolate characteristic Name Size (kb) Transformant Pulsotype Sequence type blaKPC-2 TnpA ISKPN6 ISKPN7 ECa PAb PA-1 Medellin 2006 Bronchial secretion Tertiary care center KPC-2 + + + pCOL 45 + + A1 ST308 PA-2 Bogota 2006 Stool Unknown KPC-2 − + − pPA-2 13 + + B ST1006 PA-3 Barranquiila 2006 Bronchial secretion Intensive care unit KPC-2 + + + pPA-3 60 − − C ST1060 PA-4 Medellin 2006 Bone General wardc KPC-2 + + + pPA-4 45 + + A2 ST308 PA-5 Medellin 2006 Surgical wound General ward KPC-2 + + + pPA-5 45 + + A2 ST308 PA-6 Medellin 2006 Surgical wound General ward KPC-2 + + + pPA-6 45 + + A1 ST308 PA-7 Medellin 2006 Urine General ward KPC-2 + + + pPA-7 45 + + A1 ST308 PA-8 Medellin 2006 Tracheal aspirate Intensive care unit KPC-2 + + + pPA-8 45 + + A1 ST308 PA-9 Cali 2007 Peritoneal fluid General ward KPC-2 + + + − − − − D ST235 PA-10 Pereira 2010 Urine Intensive care unit KPC-2 + + + − − − − E ST235 - Table 2.
Primers used in this study
Name of primer Fig. 2 lane no. Sequence (5′-3′) Reference KPC-A 1 CTGTCTTGTCTCTCATGGCC 11 KPC-B 2 CCTCGCTGTGCTTGTCATCC 11 3098U 3 TGACCCTGAGCGGCGAAAGC 11 4914L 4 GAAGATGCCAAGGTCAATGC 11 TnpA-178U 5 CTTGCAGATCGGCTACTTCA This study TnpA-1198U 6 CGCCATCTTCGTCCACTGTA This study TnpA-2537L 7 GCCCTGCGTCATTTCCTTCA This study ISKpn7-24U 8 CCCACCCCAGCGAAGTAAAAC This study ISKpn7-1916L 9 GACCCACTTTACCCCTGAAT This study ISKpn6-82U 10 CGTCGGCGCCAAGGATACCA This study ISKpn6-1429L 11 ATCTGCTGCCCCCTTCTCTG This study 141R-6 12 TCACCGGCCCTCACCTTTGG 11 EcoRI-out 13 CACCCGACCTGGACGAACTA 11 Delta SHVa AAGATCCACTATCGCCAGCAG 4 Delta SHVb ATTCAGTTCCGTTTCCCAGCGG 4 Pre-TEM1 GTATCCGCTCATGAGACAATA 4 Pre-TEM2 TCTAAAGTATATATGAGTAAACTTGGTCTG 4 CTX-MA CGCTTTGCGATGTGCAG 4 CTX-MB ACCGCGATATCGTTGGT 4 VIM-B ATGGTGTTTGGTCGCATATC 11 VIM-F TGGGCCATTCAGCCAGATC 1 IMP-2004A ACAYGGYTTGGTDGTTCTTG 1 IMP-2004B GGTTTAAYAAAACAACCACC 1 OXA-9A TTCGTTTCCGCCACTCTCCC 4 OXA-9B ACGAGAATATCCTCTCGTGC 4
