Use of the dnaJ Gene for the Detection and Identification of All Legionella pneumophila Serogroups and Description of the Primers Used to Detect 16S rDNA Gene Sequences of Major Members of the Genus Legionella

We sequenced about 930 bp of the dnaJ gene from 15 Legionella pneumophila serogroups and some other members of the genus Legionella. As L. pneumophila 16S rDNA sequences could not discriminate between all subspecies and serogroups, we assessed the use of dnaJ gene sequences to differentiate between Legionella subspecies as well as between L. pneumophila serogroups. A phylogenetic analysis revealed that dnaJ gene sequences were more variable between the L. pneumophila serogroups than mip gene and 16S rDNA sequences. By studying 61 strains from 41 species of the genus Legionella, as well as other genera, we established a PCR method that could amplify 285 bp of dnaJ gene from all L. pneumophila serogroups. This primer set was more sensitive than mip gene primers and was able to detect 0.25 ng of purified DNA. We also describe the 16S rDNA primers that were used to detect most Legionella genus members.

[1]  P. Christen,et al.  Mechanism of the Targeting Action of DnaJ in the DnaK Molecular Chaperone System* , 2003, Journal of Biological Chemistry.

[2]  S. Chakrabarti,et al.  Development of a new sensitive and efficient multiplex polymerase chain reaction (PCR) for identification and differentiation of different mycobacterial species , 2003, Tropical medicine & international health : TM & IH.

[3]  Jane S Tang,et al.  Characterization of Members of the Legionellaceae Family by Automated Ribotyping , 2003, Journal of Clinical Microbiology.

[4]  Hiroshi Miyamoto,et al.  Application of RNA Polymerase β-Subunit Gene (rpoB) Sequences for the Molecular Differentiation of Legionella Species , 2002, Journal of Clinical Microbiology.

[5]  L. Franzin,et al.  Legionella taurinensis, a new species of Legionella isolated in Turin, Italy. , 2000, International journal of systematic and evolutionary microbiology.

[6]  C. Saint,et al.  A PCR test for the identification and discrimination of Legionella longbeachae serogroups 1 and 2. , 1999, Journal of microbiological methods.

[7]  F. Vandenesch,et al.  Legionella taurinensis sp. nov., a new species antigenically similar to Legionella spiritensis. , 1999, International journal of systematic bacteriology.

[8]  D. Murdoch,et al.  Sequence-Based Classification Scheme for the GenusLegionella Targeting the 5S rRNA Gene , 1999, Journal of Clinical Microbiology.

[9]  F. Vandenesch,et al.  Identification of Legionella Species by Random Amplified Polymorphic DNA Profiles , 1998, Journal of Clinical Microbiology.

[10]  P. Normand,et al.  Species identification of Legionella via intergenic 16S-23S ribosomal spacer PCR analysis. , 1998, International journal of systematic bacteriology.

[11]  P. Manning,et al.  Sequence-Based Classification Scheme for the GenusLegionella Targeting the mip Gene , 1998, Journal of Clinical Microbiology.

[12]  P. Manning,et al.  Sequence Analysis of the mip Gene of the Soilborne Pathogen Legionella longbeachae , 1998, Infection and Immunity.

[13]  D. Raoult,et al.  Phylogenetic diversity of intra-amoebal legionellae as revealed by 16S rRNA gene sequence comparison. , 1996, Microbiology.

[14]  J. Pruckler,et al.  Legionella-like amebal pathogens--phylogenetic status and possible role in respiratory disease. , 1996, Emerging infectious diseases.

[15]  A. Karzai,et al.  A Bipartite Signaling Mechanism Involved in DnaJ-mediated Activation of the Escherichia coli DnaK Protein (*) , 1996, The Journal of Biological Chemistry.

[16]  P. V. van Helden,et al.  Strain-specific variation in the dnaJ gene of mycobacteria. , 1996, Journal of medical microbiology.

[17]  R. Birtles,et al.  Phylogeny of Legionellaceae based on small-subunit ribosomal DNA sequences and proposal of Legionella lytica comb. nov. for Legionella-like amoebal pathogens , 1996 .

[18]  H. Bujard,et al.  A cycle of binding and release of the DnaK, DnaJ and GrpE chaperones regulates activity of the Escherichia coli heat shock transcription factor sigma32. , 1996, The EMBO journal.

[19]  C. Georgopoulos,et al.  Both ambient temperature and the DnaK chaperone machine modulate the heat shock response in Escherichia coli by regulating the switch between sigma 70 and sigma 32 factors assembled with RNA polymerase. , 1995, The EMBO journal.

[20]  A. Macario,et al.  The ArchaealdnaK-dnaJGene Cluster: Organization and Expression in the MethanogenMethanosarcina mazei , 1995 .

[21]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[22]  T. Ezaki,et al.  Comparison of Detection Methods for Legionella Species in Environmental Water by Colony Isolation, Fluorescent Antibody Staining, and Polymerase Chain Reaction , 1993, Microbiology and immunology.

[23]  E. Conway de Macario,et al.  dnaJ in Archaea. , 1993, Nucleic acids research.

[24]  R. Breiman,et al.  Legionella: Current Status and Emerging Perspectives , 1993 .

[25]  H. Bujard,et al.  Physical interaction between heat shock proteins DnaK, DnaJ, and GrpE and the bacterial heat shock transcription factor σ 32 , 1992, Cell.

[26]  C. Georgopoulos,et al.  Biological role and regulation of the universally conserved heat shock proteins. , 1991, The Journal of biological chemistry.

[27]  S. Warwick,et al.  The use of 16S ribosomal RNA analyses to investigate the phylogeny of the family Legionellaceae. , 1991, Journal of general microbiology.

[28]  A. Reingold,et al.  Legionella pneumonia in the United States: the distribution of serogroups and species causing human illness. , 1984, The Journal of infectious diseases.

[29]  A. Steigerwalt,et al.  Classification of the Legionnaires' disease bacterium: Legionella pneumophila, genus novum, species nova, of the family Legionellaceae, familia nova. , 1979, Annals of internal medicine.

[30]  W. Dowdle,et al.  Legionnaires' disease: isolation of a bacterium and demonstration of its role in other respiratory disease. , 1977, The New England journal of medicine.

[31]  K. Ko,et al.  Legionella busanensis sp. nov., isolated from cooling tower water in Korea. , 2003, International journal of systematic and evolutionary microbiology.

[32]  F. Grimont,et al.  rRNA gene restriction patterns of Legionella species: a molecular identification system. , 1989, Research in microbiology.