Molecular epidemiology of Legionella pneumophilia serogroup 1 by ribotyping with a non-radioactive probe and PCR fingerprinting

Abstract

Hybridization with acetylaminofluorene-labelled 16 + 23 S rRNA from Escherichia coli was used to detect DNA polymorphism among Legionella pneumophila serogroup 1 isolates. Isolates from unrelated patients showed at least four different rRNA restriction patterns, whereas those from related patients showed a single pattern. Amplification of genomic regions with an arbitrary primer by polymerase chain reaction was used to further analyze the isolates. Related isolates showed closely related patterns while unrelated isolates displayed six distinct patterns. We could differentiate the majority of unrelated isolates with the combination of the patterns obtained with the ribotyping and the PCR fingerprinting, while strains from the same outbreak remained highly related. The ribotyping and the PCR fingerprinting are proposed as useful and easy to perform epidemiological markers of L. pneumophila serogroup 1 infection.

Identification and DNA fingerprinting of Legionella strains by randomly amplified polymorphic DNA analysis

“…PCR-based DNA fingerprinting techniques with single, short, random oligonucleotide primers have successfully been used to analyze genomic DNAs of viral, bacterial, fungal, plant, and animal origin (1,18). Subtyping of L. pneumophila isolates by these techniques with a single random primer has also been attempted (3,10,13,17). The discriminatory ability of the RAPD fingerprint profile may be expanded by using multiple primers in the PCR assay.…”

“…Recently PCR-based DNA fingerprinting methods such as randomly amplified polymorphic DNA (RAPD) analysis and repetitive-element PCR were introduced as reliable methods of distinguishing between bacterial species and strains. Subtyping of Legionella pneumophila isolates with single, short, random oligonucleotide primers has been reported (2,10,13,17). Most of these published methods place emphasis on the typing of L. pneumophila serogroup 1, probably because this serogroup appears to be responsible for the majority of legionellosis cases.…”

A multicenter evaluation of genotypic methods for the epidemiologic typing of Legionella pneumophila serogroup 1: results of a pan-European study

“…Many methods have been described for the genotypic and phenotypic typing of L. pneumophila sg 1 [12–32]. However, with few exceptions, these methods have been used in a single laboratory, for the simultaneous examination of a few isolates.…”

Μοριακή τυποποίηση στελεχών legionella pneumophila ορολογικής ομάδας 1 και συσχέτιση της μοριακής δομής των στελεχών με τη λοιμογόνο δύναμη τους

“…σε βακτηριοφάγο λ: λΝ8 20, λΝ8 21) (349), β) ετερόλογο ή ομόλογο ριβοσωμικό RNA (16S και 23S RNA). Η προσέγγιση αυτής της μεθόδου είναι γνωστή ως ριβοτυτάα (ribotyping) (345,350,351). Ο ανιχνευτής σημαίνεται είτε ραδιενεργά, οπότε η αξιολόγηση των αποτελεσμάτων επιτυγχάνεται με ανάπτυξη σήματος μετά από αυτοραδιογραφία, είτε μη ραδιενεργά (συνήθως με βιοτίνη ή διγοξιγενίνη), οπότε τα μοριακά πρότυπα αξιολογούνται μετά από ανάπτυξη χρώματος με ενζυμική αντίδραση (349,350).…”