title: The N-acylneuraminate cytidyltransferase gene, neuA, is heterogenous in Legionella pneumophila strains but can be used as a marker for epidemiological typing in the consensus sequence-based typing scheme.
abstract: Sequence-based typing (SBT) is the internationally recognized standard method for genotyping Legionella pneumophila. To date all strains of serogroup 1 (SG1) and some of SGs 2 to 14 yield a seven-allele profile and can be assigned a sequence type (ST). However, for some strains belonging to SGs 2 to 14, the targeted region of the neuA gene could not be amplified using the published standard primers. We determined the DNA sequence of a neuA gene homolog located in the lipopolysaccharide synthesis locus of strain Dallas-1E. By using newly designed degenerate consensus primers based on the neuA homolog in strains Dallas-1E, Philadelphia-1, Paris, Lens, and Corby, we were able to obtain DNA sequences for all 48 non-SG1 strains which were untypeable by the standard method. Our data show that the neuA gene is present in all L. pneumophila strains but differs significantly in some non-SG1 strains at both the DNA and amino acid levels. The new primers can be used to amplify and sequence the neuA gene in all strains and can substitute for the standard primers. This offers the possibility of assigning an ST to all strains of L. pneumophila.
authors.affiliation: Institut für Medizinische Mikrobiologie und Hygiene, TU Dresden, Fiedlerstrasse 42, D-01307 Dresden, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Farhat
authors.forename: Massimo
authors.initials: M
authors.lastname: Mentasti
authors.forename: Enno
authors.initials: E
authors.lastname: Jacobs
authors.forename: Norman K
authors.initials: NK
authors.lastname: Fry
authors.forename: Christian
authors.initials: C
authors.lastname: Lück
mesh_terms: D000123:Acetyltransferases / Q000235:genetics*; D016000:Cluster Analysis; D016384:Consensus Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D014644:Genetic Variation; D005838:Genotype; D016952:Legionella pneumophila / Q000145:classification* / Q000201:enzymology* / Q000235:genetics; D017720:Molecular Epidemiology / Q000379:methods; D008969:Molecular Sequence Data; D058889:Molecular Typing / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D000123:Acetyltransferases