Identification of Burkholderia species and genomovars from cystic fibrosispatients by AFLP fingerprinting

AFLP is a genomic fingerprinting technique based on the selective amplifica tion of restriction fragments from a total double-digest of genomic DNA. Th e applicability of this method to differentiate between species and genomov ars of the genus Burkholderia was tested, with particular emphasis on taxa occurring in cystic fibrosis patients. In this study, 78 well-characterized strains and field isolates were investigated by two methods of AFLP finger printing. In the manual procedure, a radioactively labelled primer was used , amplified fragments were separated by conventional PACE and the patterns were revealed by autoradiography. In the automated procedure, a fluorescent ly labelled primer was used in combination with electrophoresis and on-line data collection by means of an automated DNA sequencer. Overall, there was good agreement between the two AFLP procedures and the data were mostly co nsistent with results obtained from SDS-PAGE of whole-cell proteins and DNA -DNA hybridization experiments. The automated AFLP procedure has considerab le technical advantages compared with the manual AFLP procedure, but a thor ough visual analysis of the DNA profiles was required to avoid misidentific ation of some Burkholderia cepacia genomovar III strains.