Identification and characterization of nine atypical Candida dubliniensis clinical isolates

Albaina, Olatz; Sahand, Ismail H.; Brusca, María Isabel; Sullivan, Derek J.; Larrinoa, Iñigo Fernández de; Moragues, Marı́a D.

doi:10.1099/jmm.0.078832-0

Cited by 7 publications

(6 citation statements)

References 58 publications

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“…As a result, our assay circumvents the imperfections of phenotypic assays (CHROMagar, VITEK 2 and ID 32C) that suffer from the lack of specificity [ 38 , 39 ]. Additionally, subjecting various populations of the same species to phenotypic assay could showed various results [ 32 , 40 , 41 ]. Accordingly, as suggested by Griseo et al (2015), small clinical laboratories can take advantage of specific phenotypic methods supplemented with easy-to-perform PCR-based approaches to identify and report isolated cryptic Candia species [ 8 ].…”

Section: Discussionmentioning

confidence: 99%

Identification of nine cryptic species of Candida albicans, C. glabrata, and C. parapsilosis complexes using one-step multiplex PCR

et al. 2018

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BackgroundCandida albicans, Candida glabrata, and Candida parapsilosis are three prevalent causes of candidiasis, worldwide. These species are considered as nine medically important complex species. Limited knowledge about these newly recognized species prompted us to develop a one-step, multiplex PCR to detect and identify them in clinical settings.MethodsPrimers targeting Hyphal Wall Protein I gene for the C. albicans, C. dubliniensis, C. africana, Intergenic Spacer for the C. glabrata, C. nivariensis, C. bracarensis, and Intein and ITS rDNA for the C. parapsilosis, C. orthopsilosis, and C. metapsilosis were designed. Using 168 CBS reference strains and 280 clinical isolates, the specificity and reproducibility of the developed assay were evaluated.ResultsOur developed assay successfully identified and distinguished all the nine species. No cross-reaction with closely- and distantly-related yeast species, Aspergillus species and human DNA was observed, resulting in 100% specificity. The ambiguous results obtained by MALDI-TOF for C. albicans and C. africana were corrected by our 9-plex PCR assay. This assay identified all the cryptic complex species from two test sets from Iran and China, correctly.ConclusionsOur developed multiplex assay is accurate, specific, cost/time-saving, and works without the tedious DNA extraction steps. It could be integrated into routine clinical laboratories and as a reliable identification tool and has the potential to be implemented into epidemiological studies to broaden the limited knowledge of cryptic species complexes.Electronic supplementary materialThe online version of this article (10.1186/s12879-018-3381-5) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Identification of nine cryptic species of Candida albicans, C. glabrata, and C. parapsilosis complexes using one-step multiplex PCR

et al. 2018

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“…Presumptive identification by these methods used routinely in low-complexity laboratories continues to be a problem because they do not provide definitive identification data. Distinction between these two species is important in terms of treatment, in order to understand the clinical and epidemiological significance of the role played by C. dubliniensis in human infections 4,5,22,[24][25][26][27] .…”

Section: Discussionmentioning

confidence: 99%

gDNA extraction from Candida albicans and Candida dubliniensis in subgingival samples in Argentina. Evaluation of different methods

Dubois,

Salgado,

Gliosca

et al. 2023

Acta Odontol Latinoam

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The oral cavity constitutes a unique ecosystem with highly variable ecological niches that harbor a great variety of microorganisms, including yeasts. Molecular methods are currently considered the gold standard for identifying species, although they involve limitations associated with the disruption of yeast cell walls to release the genomic DNA (gDNA) for amplification. Aim: The aim of this study was to compare the performance of different methods for extracting gDNA from Candida albicans and Candida dubliniensis, subsequently amplifying DNA by PCR. Materials and Method: Fifty-two isolates (16 C. albicans and 36 C. dubliniensis) were obtained from subgingival biofilm of HIV+ patients with clinical signs of periodontal disease. The study evaluated 6 gDNA extraction methods and two PCR amplification methods. Furthermore, the presence of alleles of HWP1 gene was determined in C. albicans. Results: Comparisons of six methods show statistically significant differences (p<0.001) except for C. albicans in two of them. For C. dubliniensis, statistical differences were observed in all comparisons. Commercial methods were more efficient for concentrating gDNA than in-house methods, and both PCRs were effective. Ten heterozygous C. albicans isolates for this allele were positive for the HWP1-1 / HWP1-2 allele, one was homozygous for Wild Type HWP1-1 allele, and 5 were homozygous for novel/rare HWP1-2 allele. Conclusions: This study aims to provide simple, inexpensive strategies for phenotypic identification and molecular confirmation of Candida albicans and Candida dubliniensis for non-reference laboratories with low complexity and/or low budgets.

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“…Microscopic evaluation results are usually subjective and the decision on species identification depends on experience and species characteristics, and several factors such as incubation duration may influence morphological differentiation as well. Although Saeed et al [ 19 ] tested CMTA for direct inoculation of the samples, they have found that CMTA can easily differentiate common Candida species; however, identification of rare ones is still challenging [ 20 , 21 ]. The results of our study are also similar to that of the previous study.…”

Section: Discussionmentioning

confidence: 99%

Medically important Candida spp. identification: an era beyond traditional methods

EVREN¹,

GÖÇMEN²,

İŞTAR³

et al. 2022

Turkish Journal of Medical Sciences

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Background/aim Candida infections are gaining more attention for the last few decades so diagnostic tools are very important for early diagnosis. Conventional identification of yeasts is time-consuming, molecular methods are more complicated and relatively expensive gold-standard methods. Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) was put into the market due to its speed and high accuracy. The aim of this study was to evaluate the performance of corn meal tween-80 agar (CMTA), CHROMagar Candida medium, and MALDI-TOF MS and to compare the obtained results with DNA sequencing. Materials and methods The CHROMagar Candida medium, CMTA, and MALDI-TOF MS Biotyper System were used to test 416 isolates. The isolates with discrepant results by at least one of the three methods were subjected to sequence analysis. Results The identification results of the 351 (%84.4) were compatible with all three methods. When compared to the sequencing results, the most accurate results were obtained by the MALDI-TOF MS, especially for rare Candida species. Conclusion MALDI-TOF MS is found to be the most accurate identification tool for clinically important Candida strains. CMTA alone should not be used for the final identification of Candida species and the chromogenic medium should always be considered presumptive.

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Identification and characterization of nine atypical Candida dubliniensis clinical isolates

Cited by 7 publications

References 58 publications

Identification of nine cryptic species of Candida albicans, C. glabrata, and C. parapsilosis complexes using one-step multiplex PCR

Identification of nine cryptic species of Candida albicans, C. glabrata, and C. parapsilosis complexes using one-step multiplex PCR

gDNA extraction from Candida albicans and Candida dubliniensis in subgingival samples in Argentina. Evaluation of different methods

Medically important Candida spp. identification: an era beyond traditional methods

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