USE OF AMPLIFIED FRAGMENT LENGTH POLYMORPHISM IN MOLECULAR TYPING OF LEGIONELLA-PNEUMOPHILA AND APPLICATION TO EPIDEMIOLOGIC STUDIES

A novel method for molecular typing of organisms, amplified fragment l ength polymorphism analysis, was tested for its suitability in epidemi ological studies in medical microbiology, Amplified fragment length po lymorphism analysis, originally developed for typing crop plants, cons ists of a simple restriction-ligation reaction and a subsequent PCR am plification, In a single-step reaction, the genomic DNA is digested an d the restriction fragments are ligated to specially constructed adapt ers, PCR amplification of such tagged restriction fragments with prime rs complementary to the adapters allows the detection of restriction f ragment length polymorphisms upon resolution on agarose gels, The meth od is fast, efficient, and reproducible for typing strains of Legionel la pneumophila isolated from both humans and the environment, The accu racy of the method was tested by comparison with standard restriction fragment length polymorphism typing performed with both a ribosomal an d a genomic probe.