Advanced search
Start date
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Accurate Identification of Candida parapsilosis (Sensu Lato) by Use of Mitochondrial DNA and Real-Time PCR

Full text
Souza, Ana Carolina R. [1] ; Ferreira, Renata C. [2, 1, 3] ; Goncalves, Sarah S. [1] ; Quindos, Guillermo [4] ; Eraso, Elena [4] ; Bizerra, Fernando C. [1] ; Briones, Marcelo R. S. [2, 3] ; Colombo, Arnaldo L. [1]
Total Authors: 8
[1] Univ Fed Sao Paulo, Lab Especial Micol, Disciplina Infectol, Sao Paulo - Brazil
[2] Univ Fed Sao Paulo, Dept Microbiol Imunol & Parasitol, Sao Paulo - Brazil
[3] Univ Fed Sao Paulo, Lab Genom Evolut & Biocomplexidade, Sao Paulo - Brazil
[4] Univ Pais Vasco Euskal Herriko Unibertsitatea, Lab Micol Med, Dept Inmunol Microbiol & Parasitol, Fac Med & Odontol, Bilbao - Spain
Total Affiliations: 4
Document type: Journal article
Source: Journal of Clinical Microbiology; v. 50, n. 7, p. 2310-2314, JUL 2012.
Web of Science Citations: 19

Candida parapsilosis is the Candida species isolated the second most frequently from blood cultures in South America and some European countries, such as Spain. Since 2005, this species has been considered a complex of 3 closely related species: C. parapsilosis, Candida metapsilosis, and Candida orthopsilosis. Here, we describe a real-time TaqMan-MGB PCR assay, using mitochondrial DNA (mtDNA) as the target, which readily distinguishes these 3 species. We first used comparative genomics to locate syntenic regions between these 3 mitochondrial genomes and then selected NADH5 as the target for the real-time PCR assay. Probes were designed to include a combination of different single-nucleotide polymorphisms that are able to differentiate each species within the C. parapsilosis complex. This new methodology was first tested using mtDNA and then genomic DNA from 4 reference and 5 clinical strains. For assay validation, a total of 96 clinical isolates and 4 American Type Culture Collection (ATCC) isolates previously identified by internal transcribed spacer (ITS) ribosomal DNA (rDNA) sequencing were tested. Real-time PCR using genomic DNA was able to differentiate the 3 species with 100% accuracy. No amplification was observed when DNA from other species was used as the template. We observed 100% congruence with ITS rDNA sequencing identification, including for 30 strains used in blind testing. This novel method allows a quick and accurate intracomplex identification of C. parapsilosis and saves time compared with sequencing, which so far has been considered the ``gold standard{''} for Candida yeast identification. In addition, this assay provides a useful tool for epidemiological and clinical studies of these emergent species. (AU)

FAPESP's process: 07/08575-1 - Hematogenic candidiasis: a multidisciplinary approach for the improvement of diagnostic tools and caracterization of biomarkers related to its prognosis
Grantee:Arnaldo Lopes Colombo
Support type: Research Projects - Thematic Grants
FAPESP's process: 09/01230-4 - Clinical aplications on the molecular systematic of Candida spp. in the perspective of comparative genomics of gene genealogies and Bayesian analyses
Grantee:Renata Carmona e Ferreira
Support type: Scholarships in Brazil - Post-Doctorate
FAPESP's process: 10/17179-5 - Proteomics as a strategy to identify biomarkers for early diagnosis of candidemia
Grantee:Fernando César Bizerra
Support type: Scholarships in Brazil - Post-Doctorate
FAPESP's process: 10/02752-1 - Identification of species from the Candida "para-meta-ortho-psilosis" complex using real-time PCR
Grantee:Ana Carolina Remondi Souza
Support type: Scholarships in Brazil - Master