Application of Amplified Fragment Length Polymorphism Analysis to Subtyping of Legionella pneumophila Serogroup 6

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Abstract
One patient isolate and one environmental isolate of Legionella pneumophila serogroup 6 were obtained from an outbreak investigation associated with a whirlpool spa, in which 2 cases of Legionnaires' disease and 10 cases of Pontiac fever were diagnosed. The authors used amplified fragment length polymorphism (AFLP) analysis to type the patient and environmental isolates. They compared the patterns obtained from the outbreak-related strains with the electrophoretic types obtained from 48 strains analyzed by multilocus enzyme electrophoreis (MLEE). The previous analysis by MLEE had grouped the 48 unrelated strains of serogroup 6 in to 11 electrophoretic types. L. pneumophila serogroup 6 strains Chicago 2, Johannesburg 5, Albany 1, Oxford 1, SRP 39, Denver 3, Sydney 1, LD 82-683, ED 38, and Vasteras 57/1, the whirlpool and patient isolates, were selected for evaluation of the AFLP technique on the basis of previous subtyping performed by MLEE. Analysis of fragment sizes by agarose gel electrophoresis indicated that the patient and environmental strains were identical; however the 10 unique strains were divided into 7 patterns. By incorporating a fluorescent primer and separating the fragments with the ABI 310, all 10 unrelated strains could be shown to have unique patterns that were separate from the patient and environmental strains. An advantage of AFLP compared with pulsed-field gel electrophoresis or MLEE is that it is easier to perform and less time-consuming.

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