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J Med Microbiol 54 (2005), 333-346; DOI: 10.1099/jmm.0.45690-0
© 2005 Society for General Microbiology
ISSN 0022-2615

Amplified fragment length polymorphism (AFLP) versus randomly amplified polymorphic DNA (RAPD) as new tools for inter- and intra-species differentiation within Bordetella

Anna Gzyl1, Ewa Augustynowicz1, Ewa Mosiej2, Monika Zawadka1, Grzegorz Gniadek1, Aneta Nowaczek1 and Janusz Slusarczyk1

1Department of Sera and Vaccine Evaluation, National Institute of Hygiene, 24 Chocimska Str., 00-791 Warsaw, Poland 2Interfaculty Studies of Biotechnology, Warsaw Agricultural University, 159 Nowoursynowska Str., 00-776 Warsaw, Poland

Correspondence Anna Gzyl agzyl{at}pzh.gov.pl

Received April 5, 2004
Accepted November 19, 2004

Automated amplified fragment length polymorphism (AFLP) and randomly amplified polymorphic DNA (RAPD) techniques with fluorescently labelled primers were used to track differences among isolates of the eight known species of the Bordetella genus. Eighty-one representative strains of these species from international and Polish bacterial collections were genotyped according to RAPD protocols using primer 1254 or 1247, and AFLP involving EcoRI/MseI or newly designed SpeI/ApaI restriction/ligation/amplification procedures. By comparing AFLP and RAPD data, it was concluded that the discriminatory power of AFLP is higher in comparison with RAPD for both intra- and inter-species differentiation of isolates of the Bordetella genus. The most precise level of inter-species discrimination and the highest level of intra-species discrimination of the Bordetella isolates of the eight species were observed in the AFLP EcoRI/MseI and SpeI/ApaI sets, respectively. Both techniques might provide alternative tools for the identification of Bordetella at the genomic species and strain levels, and thus may be valuable in human and veterinary diagnostics as well as in epidemiology. By applying the AFLP technique presented in this article, more precise data on the emergence of newly acquired and/or on expanded clones and transmission routes of isolates of the Bordetella genus in the human and animal environments might be obtained.

Abbreviations: AFLP, amplified fragment length polymorphism; IS, insertion sequence; MLEE, multilocus enzyme electrophoresis; MLST, multilocus sequence typing; RAPD, randomly amplified polymorphic DNA; REA, restriction enzyme analysis; UPGMA, unweighted pair group method using arithmetic averages.






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