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Review
. 1999 Oct;37(10):3083-91.
doi: 10.1128/JCM.37.10.3083-3091.1999.

Amplified-fragment length polymorphism analysis: the state of an art

P H Savelkoul  1 , H J Aarts, J de Haas, L Dijkshoorn, B Duim, M Otsen, J L Rademaker, L Schouls, J A Lenstra
Affiliations
Review

Amplified-fragment length polymorphism analysis: the state of an art

P H Savelkoul et al. J Clin Microbiol. 1999 Oct.
No abstract available

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Figures

FIG. 1
FIG. 1
Schematic representation of the AFLP analysis principle. 1, point mutations incorporated in the adapter sequences to prevent digestion after ligation are shaded. 2, one of the primers is labeled. In this representation both primers contain one selective nucleotide (shaded) in the unknown fragment.
FIG. 2
FIG. 2
Example of fluorescently labeled AFLP patterns and dendrogram for 11 different Klebsiella isolates. Patterns are the result of amplification of templates generated after restriction and ligation as shown in Fig. 1. The fragments were analyzed on an automated Vistra sequencer (Amersham-Pharmacia Biotech). The dendrogram was constructed with GelCompar (Applied Maths) software by using the Pearson correlation and cluster analysis by the unweighted pair group method using arithmetic averages. Percentages of similarity and molecular sizes (in base pairs) are shown above the dendrogram. Lanes 1 to 8, identical Klebsiella pneumoniae isolates; lanes 9 and 10, different K. pneumoniae strains; lane 11, a Klebsiella oxytoca strain. Within the AFLP patterns from Klebsiella, for instance, three windows of similarity may be applicable on the basis of the described experimental conditions: window I, 90 to 100% homology, identical strains; window II, 60 to 90% homology, different strains, same species (e.g., Klebsiella pneumoniae); window III, 40 to 60% homology, different species of the same genus; window IV, less than 40% homology, species from different genera.
FIG. 3
FIG. 3
Relative applicability of various fingerprinting and DNA techniques at different levels of taxonomic resolution. Reprinted from reference with permission of the publishers. tRNA-PCR, tRNA intergenic spacer region PCR; ITS-PCR, 16S-23S rRNA intergenic spacer region PCR; LFRA, low-frequency restriction enzyme analysis; AP-PCR, arbitrarily primed PCR. The other abbreviations are defined in the text.
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