Use of PCR-restriction enzyme pattern analysis and sequencing database for hsp65 gene-based identification of Nocardia species.
Nocardia identification required laborious and time-consuming phenotypic and chemotaxonomic methods until molecular methods were developed in the mid-1990s. Here we reassessed the capacity of PCR-restriction enzyme pattern analysis (PRA) of the hsp65 gene to differentiate Nocardia species, including 36 new species. Our results confirm that hsp65 PRA must no longer be used for Nocardia species identification, as many species have the same restriction pattern. We then compared sequencing-based strategies using an hsp65 database and a 16S rRNA database and found that the hsp65 region contained sufficient polymorphisms for comprehensive Nocardia species identification.
Rodríguez-Nava
Verónica
V
UMR CNRS 5557, Laboratoire de Mycologie Fondamentale et Appliquée aux Biotechnologies Industrielles, UFR Pharmacie, UCB Lyon I, 69373, Lyon Cedex 08, France.
Couble
Andrée
A
Devulder
Gregory
G
Flandrois
Jean-Pierre
JP
Boiron
Patrick
P
Laurent
Frédéric
F
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D030541:Databases, Genetic; D020459:Genes, rRNA; D006360:Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D009615:Nocardia / Q000145:classification* / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D001426:Bacterial Proteins; D006360:Heat-Shock Proteins; D012336:RNA, Ribosomal, 16S