M021: Differentiation of Burkholderia (Pseudomonas) Gladioli
and Clinical and Environmental Strains of Burkholderia
Cepacia by Multiple Arbitrary Amplicon Profiling (MAAP).
J.L. Tipper., E. Ingham, J.H. Cove, N.J. Todd1, J.M. Littlewood2,
D.E. Stead3 & K.G. Kerr (Dept. of Microbiology, University
of Leeds, Depts. of 1Microbiology & 2Paediatrics, St. James's
University Hospital, Leeds. 3Central Science Laboratory, M.A.F.F.,
Harpenden, UK).
Colonisation of cystic fibrosis (CF) patients by B. cepacia often results in asymptomatic carriage of this organism, however other patients experience rapid clinical deterioration. Burkholderia gladioli is difficult to differentiate from B. cepacia by biochemical tests and recent reports suggest that clinical isolates from CF patients possess characteristics of both B. cepacia and B. gladioli, and therefore the isolation of both organisms should be regarded with equal clinical significance. MAAP has been used to discriminate between virulent and non-virulent micro-organisms and this study aimed to determine whether MAAP was a suitable method for differentiating strains of B. gladioli and B. cepacia from clinical and environmental sources. Total genomic DNA was extracted from all strains and MAAP was performed using the following 17 bp random primer 5'- GCCCCCAGGGGCACAGT -3'. The reaction products were analysed by agarose gel electrophoresis. Data obtained from a small number of well characterised strains suggests that MAAP is capable of differentiating between B. gladioli and B. cepacia and also between clinical and environmental isolates of B. cepacia MAAP may represent a rapid and reproducible method for differentiating strains of B. cepacia and B. gladioli and further studies with a larger number of strains are warranted. In addition, we intend to use this method to correlate clinical status of patients colonised with B. cepacia with MAAP type, and the ability of isolates to invade respiratory epithelial cells, which may represent an important virulence factor for B. cepacia.