Efficacy of practised screening methods for detection of cephalosporin-resistant Enterobacteriaceae.

OBJECTIVES: Enterobacteriaceae with extended-spectrum beta-lactamases (ESBLs) are now widespread and simple phenotypic tests are required to detect them in diagnostic laboratories. We investigated the performance of screening methods at 16 hospitals in South-East England. METHODS: Sixteen laboratories in South-East England submitted 1195 consecutive Enterobacteriaceae isolates found to be resistant, by their routine methods, to any or all of cefpodoxime, ceftazidime and cefotaxime. These isolates were re-tested centrally with various cephalosporin/clavulanate combinations and with multiplex PCR for bla(CTX-M) and bla(AmpC) alleles. RESULTS: Screening methods among the laboratories were the following: cefpodoxime discs alone (1 site); cefpodoxime, cefotaxime and ceftazidime discs (9 sites) or agar dilution (1 site); Phoenix (2 sites), Vitek 1 (1 site) and Vitek 2 (2 sites). A total of 8% of isolates submitted based on disc tests proved fully cephalosporin-susceptible, compared with 3% sent based on tests with automated systems and none of those sent based on agar dilution tests. Among isolates submitted solely on cefpodoxime resistance 256/372 (69%) proved cephalosporin-susceptible or had only borderline resistance with no clear mechanism demonstrable; this proportion decreased to 28/160 (18%) for those submitted on the basis of resistance to ceftazidime, 18/122 (15%) for those resistant to cefotaxime and 26/496 (5%) for those resistant to both cefotaxime and ceftazidime. The inference of ESBL production by Vitek 2 had the best agreement with reference laboratory results. CONCLUSIONS: Many isolates found resistant only to cefpodoxime at the source sites proved not to have ESBLs or AmpC; screening with cefotaxime and ceftazidime allowed better specificity for identification of mechanism-based resistance, as did the automated systems. Cefpodoxime disc tests nevertheless remain a useful primary screen for laboratories prepared only to test one agent.

Community and hospital spread of Escherichia coli producing CTX-M extended-spectrum beta-lactamases in the UK.

OBJECTIVES: During 2003, the Health Protection Agency's Antibiotic Resistance Monitoring and Reference Laboratory began to receive isolates of Escherichia coli for confirmation of extended-spectrum beta-lactamase production with a phenotype implying a CTX-M-type beta-lactamase, i.e. MICs of cefotaxime > or = 8-fold higher than MICs of ceftazidime. Many were referred as being from community patients. We examined 291 CTX-M-producing isolates from the UK and investigated the genetic basis of their phenotype. METHODS: PCR was used to detect alleles encoding CTX-M enzymes and to assign these to their blaCTX-M phylogenetic groups. Selected alleles were sequenced. Producers were compared by analysis of banding patterns generated by pulsed-field gel electrophoresis of XbaI-digested genomic DNA. MICs were determined by an agar dilution method or by Etest. RESULTS: Of 291 CTX-M-producing E. coli isolates studied from 42 UK centres, 70 (24%) were reportedly from community patients, many of whom had only limited recent hospital contact. Community isolates were referred by 12 centres. Two hundred and seventy-nine (95.9%) producers contained genes encoding group 1 CTX-M enzymes and 12 contained blaCTX-M-9-like alleles. An epidemic CTX-M-15-producing strain was identified, with 110 community and inpatient isolates referred from six centres. Representatives of four other major strains also produced CTX-M-15, as did several sporadic isolates examined. Most producers were multi-resistant to fluoroquinolones, trimethoprim, tetracycline and aminoglycosides as well as to non-carbapenem beta-lactams. CONCLUSIONS: CTX-M-producing E. coli are a rapidly developing problem in the UK, with CTX-M-15 particularly common. The diversity of producers and geographical scatter of referring laboratories indicates wide dissemination of blaCTX-M genes. Because of the public health implications, including for the treatment of community-acquired urinary tract infections, the spread of these strains--and CTX-M-15 beta-lactamase in particular--merits close monitoring.