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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
Author(s) -
Liu Hongsheng,
Li Ying,
Huang Xinxiang,
Kawamura Yoshiaki,
Ezaki Takayuki
Publication year - 2003
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2003.tb03452.x
Subject(s) - biology , legionella pneumophila , legionella , primer (cosmetics) , 16s ribosomal rna , gene , phylogenetic tree , microbiology and biotechnology , polymerase chain reaction , genetics , subspecies , ribosomal dna , bacteria , paleontology , chemistry , organic chemistry
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.