Characterization of plasmids encoding extended-spectrum β-lactamases and their addiction systems circulating among Escherichia coli clinical isolates in the UK.
OBJECTIVE
To characterize plasmids encoding extended-spectrum β-lactamases (ESBLs) from a recent UK collection of clinical Escherichia coli isolates.


METHODS
The isolates comprised 118 ESBL producers referred from 54 laboratories. Plasmids were transferred by electroporation, and their incompatibility groups, associated addiction systems and resistance genes with the flanking genetic environments were identified by PCR or sequencing.


RESULTS
Seventy isolates had plasmids encoding CTX-M-15 (n = 53), CTX-M-14 (n =9), CTX-M-27 (n = 1), CTX-M-3 (n = 2) and SHV-12 (n = 5) ESBLs that were transformable; non-transformable ESBLs were mainly CTX-M enzymes (42/48). Most transformable bla(CTX-M-15) genes (43/53) were harboured on single replicon or multireplicon IncF plasmids, with IncFIA4-FIB1-FII31 (n = 11) and IncFIA1-FII2 (n = 15) being most frequent; the latter included eight pEK499 plasmids, typical of UK epidemic strain A. Plasmids harbouring bla(CTX-M-14) belonged variously to IncF, IncI1 and IncHI2 types, and 16 encoding CTX-M or SHV enzymes were non-typeable. Only IncF plasmid types carried the addiction systems sought and those with bla(CTX-M-15) frequently harboured bla(OXA-1) and aac(6')-Ib-cr, and often transferred trimethoprim and tetracycline resistance; those with bla(CTX-M-14) encoded trimethoprim, sulphonamide, streptomycin and tetracycline resistance. Most ESBL genes were associated with the well-known mobile elements ISEcp1 and IS26, but nearly half (23/55) of the ISEcp1 sequences upstream of bla(CTX-M-15) were interrupted by an IS26 at various positions.


CONCLUSIONS
Most ESBLs (70/118) were encoded by transformable plasmids, although a sizable minority could not be transformed. The majority of transformable plasmids (51/70; 72.9%) were diverse multiresistant IncF types possessing multiple addiction systems. The spread of bla(CTX-M-15) can be attributed not just to clonal expansion, but also to the horizontal dissemination of related plasmids.
Antibiotic Resistance Monitoring & Reference Laboratory, HPA-Colindale, 61 Colindale Avenue, London NW9 5EQ, UK. michel.doumith@hpa.org.uk
Michel
M
Doumith
Hiran
H
Dhanji
Matthew J
MJ
Ellington
Peter
P
Hawkey
Neil
N
Woodford
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D018274:Electroporation; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification*; D004927:Escherichia coli Infections / Q000382:microbiology*; D018014:Gene Transfer Techniques; D022761:Gene Transfer, Horizontal; D006801:Humans; D008826:Microbial Sensitivity Tests; D010957:Plasmids / Q000032:analysis*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014169:Transformation, Bacterial; D006113:United Kingdom; D001618:beta-Lactamases / Q000235:genetics*
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D001618:beta-Lactamases