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PMID: 16000805 Published · ppublish English Evaluation Study Journal Article Research Support, Non-U.S. Gov't

Development of a recA gene-based identification approach for the entire Burkholderia genus.

Applied and environmental microbiology ·Vol. 71 ·No. 7 ·2005-07-00 ·Pages 3917-27

Payne GW, Vandamme P, Morgan SH, Lipuma JJ, Coenye T, Weightman AJ, Jones TH, Mahenthiralingam E

Abstract

Burkholderia is an important bacterial genus containing species of ecological, biotechnological, and pathogenic interest. With their taxonomy undergoing constant revision and the phenotypic similarity of several species, correct identification of Burkholderia is difficult. A genetic scheme based on the recA gene has greatly enhanced the identification of Burkholderia cepacia complex species. However, the PCR developed for the latter approach was limited by its specificity for the complex. By alignment of existing and novel Burkholderia recA sequences, we designed new PCR primers and evaluated their specificity by testing a representative panel of Burkholderia strains. PCR followed by restriction fragment length polymorphism analysis of an 869-bp portion of the Burkholderia recA gene was not sufficiently discriminatory. Nucleotide sequencing followed by phylogenetic analysis of this recA fragment differentiated both putative and known Burkholderia species and all members of the B. cepacia complex. In addition, it enabled the design of a Burkholderia genus-specific recA PCR that produced a 385-bp amplicon, the sequence of which was also able to discriminate all species examined. Phylogenetic analysis of 188 novel recA genes enabled clarification of the taxonomic position of several important Burkholderia strains and revealed the presence of four novel B. cepacia complex recA lineages. Although the recA phylogeny could not be used as a means to differentiate B. cepacia complex strains recovered from clinical infection versus the natural environment, it did facilitate the identification of clonal strain types of B. cepacia, B. stabilis, and B. ambifaria capable of residing in both niches.

MeSH Terms
Bacterial Typing Techniques Burkholderia/classification,genetics Burkholderia Infections/microbiology DNA Primers DNA, Bacterial/analysis Environmental Microbiology Humans Molecular Sequence Data Phylogeny Polymerase Chain Reaction Polymorphism, Restriction Fragment Length Rec A Recombinases/genetics Sequence Analysis, DNA Species Specificity
Chemicals
DNA Primers DNA, Bacterial Rec A Recombinases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Payne George W
Cardiff School of Biosciences, Main Building, Museum Avenue, PO Box 915, Cardiff University, Cardiff, Wales CF10 3TL, United Kingdom.
Vandamme Peter
Morgan Sara H
Lipuma John J
Coenye Tom
Weightman Andrew J
Jones T Hefin
Mahenthiralingam Eshwar
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.71.7.3917-3927.2005
Published
2005-07-00
Pages
3917-27
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC1169057
Subset
IM
Databases
GENBANK
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External Links
PubMed source DOI: 10.1128/AEM.71.7.3917-3927.2005 PMC Full Text View journal details
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