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ASIARESIST Report Summary

Project ID: ICA4-CT-2001-10028
Funded under: FP5-INCO 2
Country: United Kingdom

Plate-based conjugation assay for enterobacterial isolates

A plate-based conjugation assay was optimised using a known donor isolate, E. coli RC85 (chloramphenicol resistant) and a known recipient E. coli isolate HB101 (streptomycin resistant).

Chloramphenicol resistant project isolates shown to be susceptible (> 6 mm zone size) to streptomycin by disc diffusion could be included as donor (test) isolates with the recipient isolate E. coli isolate HB101. Transconjugants were confirmed by a series of phenotypic (appearance on MacConkey agar, antibiotic resistance profiling) and genotypic tests (plasmid content and rep-PCR typing) to exclude the possibility of mutant donors or recipients.

Transconjugants typically originated from donors belonging to the family Enterobacteriaceae and involved the transfer of resistance of sulphonamide, tetracycline as well as chloramphenicol. Plasmid profiling of the transconjugants revealed that large plasmids (> 100 kb) were commonly associated with the transfer of resistance to the recipient. The conjugation assay was shown to be useful in the screening of resistance gene transfer occurring within Enterobacteriaceae isolates.

Reported by

University of Stirling
Institute of Aquaculture, Pathfoot Building, University of Stirling
FK9 4LA Stirling
United Kingdom
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