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Molecular identification of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila.

Mota AJ, Back-Brito GN, Nobrega FG - Genet. Mol. Biol. (2011)

Bottom Line: Traditional phenotypic methods and commercial kits based on carbohydrate assimilation patterns are unable to consistently distinguish among isolates of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila.As result, these species are often misidentified.Our assay was validated by DNA sequencing of the polymorphic region used in a real-time PCR assay driven by species-specific probes targeted to the fungal ITS 1 region.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Biociências, Universidade de São Paulo, São Paulo, SP, Brazil.

ABSTRACT
Traditional phenotypic methods and commercial kits based on carbohydrate assimilation patterns are unable to consistently distinguish among isolates of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila. As result, these species are often misidentified. In this work, we established a reliable method for the identification/differentiation of these species. Our assay was validated by DNA sequencing of the polymorphic region used in a real-time PCR assay driven by species-specific probes targeted to the fungal ITS 1 region. This assay provides a new tool for pathogen identification and for epidemiological, drug resistance and virulence studies of these organisms.

No MeSH data available.


Related in: MedlinePlus

Plots showing real-time PCR amplification of singleplex assays monitored with fluorescent TaqMan® MGB probes. Delta Rn versus Cycle amplification plots were obtained using probes for (A) P. guilliermondii, (B) C. palmioleophila and (C) D. hansenii templates. The NTC (no template control) curves show no detection in the absence of templates and in the presence of non-specific targets, i.e., C. palmioleophila and D. hansenii in (A), P. guilliermondii and D. hansenii in (B) and P. guilliermondii and C. palmioleophila in (C).
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f2-gmb-35-1-122: Plots showing real-time PCR amplification of singleplex assays monitored with fluorescent TaqMan® MGB probes. Delta Rn versus Cycle amplification plots were obtained using probes for (A) P. guilliermondii, (B) C. palmioleophila and (C) D. hansenii templates. The NTC (no template control) curves show no detection in the absence of templates and in the presence of non-specific targets, i.e., C. palmioleophila and D. hansenii in (A), P. guilliermondii and D. hansenii in (B) and P. guilliermondii and C. palmioleophila in (C).

Mentions: Figure 2 shows the amplification plot of the real-time PCR genotyping done using species-specific probes. The region indicated as NTC (no template control) confirmed the specificity of each probe since only in the presence of the specific target was there amplification. To confirm this finding, we analyzed the real-time PCR products in a 2% agarose gel stained with ethidium bromide and detected the expected amplicons (data not shown).


Molecular identification of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila.

Mota AJ, Back-Brito GN, Nobrega FG - Genet. Mol. Biol. (2011)

Plots showing real-time PCR amplification of singleplex assays monitored with fluorescent TaqMan® MGB probes. Delta Rn versus Cycle amplification plots were obtained using probes for (A) P. guilliermondii, (B) C. palmioleophila and (C) D. hansenii templates. The NTC (no template control) curves show no detection in the absence of templates and in the presence of non-specific targets, i.e., C. palmioleophila and D. hansenii in (A), P. guilliermondii and D. hansenii in (B) and P. guilliermondii and C. palmioleophila in (C).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3313500&req=5

f2-gmb-35-1-122: Plots showing real-time PCR amplification of singleplex assays monitored with fluorescent TaqMan® MGB probes. Delta Rn versus Cycle amplification plots were obtained using probes for (A) P. guilliermondii, (B) C. palmioleophila and (C) D. hansenii templates. The NTC (no template control) curves show no detection in the absence of templates and in the presence of non-specific targets, i.e., C. palmioleophila and D. hansenii in (A), P. guilliermondii and D. hansenii in (B) and P. guilliermondii and C. palmioleophila in (C).
Mentions: Figure 2 shows the amplification plot of the real-time PCR genotyping done using species-specific probes. The region indicated as NTC (no template control) confirmed the specificity of each probe since only in the presence of the specific target was there amplification. To confirm this finding, we analyzed the real-time PCR products in a 2% agarose gel stained with ethidium bromide and detected the expected amplicons (data not shown).

Bottom Line: Traditional phenotypic methods and commercial kits based on carbohydrate assimilation patterns are unable to consistently distinguish among isolates of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila.As result, these species are often misidentified.Our assay was validated by DNA sequencing of the polymorphic region used in a real-time PCR assay driven by species-specific probes targeted to the fungal ITS 1 region.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Biociências, Universidade de São Paulo, São Paulo, SP, Brazil.

ABSTRACT
Traditional phenotypic methods and commercial kits based on carbohydrate assimilation patterns are unable to consistently distinguish among isolates of Pichia guilliermondii, Debaryomyces hansenii and Candida palmioleophila. As result, these species are often misidentified. In this work, we established a reliable method for the identification/differentiation of these species. Our assay was validated by DNA sequencing of the polymorphic region used in a real-time PCR assay driven by species-specific probes targeted to the fungal ITS 1 region. This assay provides a new tool for pathogen identification and for epidemiological, drug resistance and virulence studies of these organisms.

No MeSH data available.


Related in: MedlinePlus