Identification of pathogenic Nocardia species by reverse line blot hybridization targeting the 16S rRNA and 16S-23S rRNA gene spacer regions.

Although 16S rRNA gene sequence analysis is employed most often for the definitive identification of Nocardia species, alternate molecular methods and polymorphisms in other gene targets have also enabled species determinations. We evaluated a combined Nocardia PCR-based reverse line blot (RLB) hybridization assay based on 16S and 16S-23S rRNA gene spacer region polymorphisms to identify 12 American Type Culture Collection and 123 clinical Nocardia isolates representing 14 species; results were compared with results from 16S rRNA gene sequencing. Thirteen 16S rRNA gene-based (two group-specific and 11 species-specific) and five 16S-23S spacer-targeted (two taxon-specific and three species-specific) probes were utilized.

Assignment of reference 5'-end 16S rDNA sequences and species-specific sequence polymorphisms improves species identification of Nocardia.

16S rDNA sequence analysis is the most accurate method for definitive species identification of nocardiae. However, conflicting results can be found due to sequence errors in gene databases. This study tested the feasibility of species identification of Nocardia by partial (5'-end 606-bp) 16S rDNA sequencing, based on sequence comparison with "reference" sequences of well-annotated strains.

Colony multiplex-tandem PCR for rapid, accurate identification of fungal cultures.

We developed a multiplex tandem PCR (MT-PCR) assay for the rapid identification of 16 fungi directly from culture. MT-PCR results were concordant with phenotypic identification for all cultures studied (n = 183). The colony MT-PCR assay was rapid (<2 h), sensitive, and specific in identifying fungal pathogens directly from primary isolation plates.

Early use of posaconazole in the successful treatment of rhino-orbital mucormycosis caused by Rhizopus oryzae.

Mucormycosis is a potentially fatal, rapidly destructive, opportunistic infection often seen in immunocompromsied individuals and patients with diabetes mellitus. We describe two patients with rhino-orbital mucormycosis caused by Rhizopus oryzae who received posaconazole within 7 days of diagnosis and in whom cure was achieved in conjunction with extensive surgical resection of diseased tissue. Posaconazole was well-tolerated.

In vitro activities of miltefosine and two novel antifungal biscationic salts against a panel of 77 dermatophytes.

The susceptibilities of 77 dermatophytes to miltefosine (MI), 1,12-bis(4-pentylpyridinium)dodecane (PYR), 1,12-bis(tributylammonium)dodecane (AM), itraconazole (ITC), terbinafine (TRB), and butenafine (BTF) were compared. Geometric mean MICs of TRB, BTF, ITC, MI, PYR, and AM were 0.039, 0.

Rapid identification of Candida species by using nuclear magnetic resonance spectroscopy and a statistical classification strategy.

Nuclear magnetic resonance (NMR) spectra were acquired from suspensions of clinically important yeast species of the genus Candida to characterize the relationship between metabolite profiles and species identification. Major metabolites were identified by using two-dimensional correlation NMR spectroscopy. One-dimensional proton NMR spectra were analyzed by using a staged statistical classification strategy.