JCM Accepts, published online ahead of print on 24 October 2007

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J. Clin. Microbiol. doi:10.1128/JCM.00906-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Genetic diversity of Actinobacillus pleuropneumoniae assessed by Amplified Fragment Length Polymorphism (AFLP) analysis

Branko Kokotovic^* and ystein Angen

National Veterinary Institute, Technical University of Denmark Bülowsvej 27, 1790 Copenhagen, Denmark

^* To whom correspondence should be addressed. Email: bko{at}vet.dtu.dk.

Amplified fragment length polymorphism (AFLP) was evaluated as a method for genotypic characterization and subtyping within the bacterial species Actinobacillus pleuropneumoniae. A total of 155 isolates of A. pleuropneumoniae, representing the serotypic variation described within this species, were analysed. In order to elucidate the species boundaries, six strains of the phylogenetically closely related species A. lignieresii were also included. Furthermore, the ability of AFLP for subtyping were studied using 42 isolates of serovar 2 and the performance compared to that obtained by PFGE.

AFLP analysis provided a clear separation of A. lignieresii and A. pleuropneumoniae and divided the isolates of A. pleuropneumoniae into 20 clusters. Most of the serovars of A. pleuropneumoniae were represented by single and quite homogeneous clusters. The exceptions were serovars 10, K2:O7 and K1:O7, which were represented by two clusters each. In the cases where the serovars were represented by more than one cluster, the existence of these clusters was supported by additional phenotypic or genotypic properties. Furthermore, AFLP-typing was able to allocate serologically nontypeable isolates to appropriate genetic groups within the species. Further investigations are needed to determine whether some of the clusters revealed through the AFLP analysis represent additional serovars. When evaluated as a subtyping method within serovar 2 of A. pleuropneumoniae, AFLP could give a separation among isolates superior to that obtained by PFGE. However, a higher degree of separation between serovar 2 isolates could be obtained by a combination of the two methods.

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