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Translation elongation factor 1-alpha gene as a marker for diagnosing of candidal onychomycosis.

ABSTRACT: Background and Purpose:Culture-based identification methods have been the gold standard for the diagnosis of candidal onychomycosis. Molecular technologies, such as polymerase chain reaction (PCR) assays, can provide an alternative for the rapid detection of Candida species. The present study was conducted to investigate a pan-Candida PCR assay based on the translation elongation factor 1-alpha (TEF-1?) gene for the detection of the most prevalent pathogenic Candida species. Materials and Methods:For the purpose of the study, an optimized pan-Candida PCR primer pair was designed, and the target was amplified and sequenced. The analytical and clinical diagnostic performance of the designed primers was tested using 17 reference strains, 137 nail scrapings suspected of onychomycosis, and 10 healthy nail specimens. Results:The use of the universal pan-Candida primers designed on TEF-1? gene resulted in the successful amplification of a 270-base pair fragment in all Candida species tested, except for C. glabrata, and reacted neither with other fungi nor with E. coli. The sequence difference count matrix showed poor insertion/deletion differences (0-2 nt) among Candida species. Among 137 nail specimens, 35% (n=48), 30.7% (n=42), and 40.1% (n=55) of the samples were found to be positive by direct microscopy, culture, and pan-Candida PCR, respectively. Conclusion:Based on the findings, the PCR-based detection targeting the DNA TEF-1? gene is a rapid and simple procedure for the diagnosis of candidal onychomycosis directly from nail sample.

SUBMITTER: Pakshir K

PROVIDER: S-EPMC7217255 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Translation elongation factor 1-alpha gene as a marker for diagnosing of candidal onychomycosis.

Pakshir Keyvan K Farazmand Forough F Ghasemi Farnoush F Mirhendi Hossein H Zomorodian Kamiar K Kharazi Mahboobeh M Alborzi Pour Ramtin R Golestani Hajar H Motamedi Marjan M

Current medical mycology 20200101 1

<h4>Background and purpose</h4>Culture-based identification methods have been the gold standard for the diagnosis of candidal onychomycosis. Molecular technologies, such as polymerase chain reaction (PCR) assays, can provide an alternative for the rapid detection of <i>Candida</i> species. The present study was conducted to investigate a pan-<i>Candida</i> PCR assay based on the translation elongation factor 1-alpha (<i>TEF-1α</i>) gene for the detection of the most prevalent pathogenic <i>Candi ...[more]

PMID: 32420503

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Project description:BackgroundRegulation of gene expression in trypanosomatids is mainly posttranscriptional. Tight regulation of mRNA stability and access to polysomes allows Trypanosoma cruzi to adapt to different environmental conditions during its life cycle. Posttranscriptional regulation requires association between mRNAs and specific proteins to form mRNP complexes. Proteins that lack a canonical RNA-binding domain, such as eukaryotic elongation factor-1α (EF-1α), may also associate with mRNPs. EF-1α is conserved in many organisms, and it plays roles in many cellular processes other than translation, including RNA transport, the cell cycle, and apoptosis.ResultsIn a previous study, EF-1α was found associated with mRNP-forming mRNAs in polysome-free fractions both in epimastigotes growing under normal conditions and in nutritionally stressed parasites. This finding suggested the possibility that EF-1α has a non-canonical function. Thus, we investigated the dynamics of EF-1α in association with T. cruzi epimastigote mRNAs under normal and stressed nutritional conditions. EF-1α is expressed throughout the parasite life cycle, but it shows a slight decrease in protein levels in the metacyclic trypomastigote form. The protein is cytoplasmically localized with a granular pattern in all forms analyzed. Following puromycin treatment, EF-1α migrated with the heaviest gradient fractions in a sucrose polysome profile, indicating that its association with large protein complexes was independent of the translation machinery. We next characterized the EF-1α-associated mRNAs in unstressed and stressed epimastigotes. We observed that specific subsets of mRNAs were associated with EF-1α-mRNPs in unstressed or stressed epimastigotes. Some mRNAs were identified in both physiological conditions, whereas others were condition-specific. Gene ontology analysis identified enrichment of gene sets involved in single-organism metabolic processes, amino acid metabolic processes, ATP and metal ion binding, glycolysis, glutamine metabolic processes, and cobalt and iron ion binding.ConclusionThese results indicate that in T. cruzi, as in other eukaryotes, EF-1α may play a non-canonical cellular role. We observed the enrichment of functionally related transcripts bound to EF-1α in normal growth conditions as well as in nutritionally stressed cell indicating a potential role of EF-1α mRNP in stress response.

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Translation elongation factor 1-alpha gene as a marker for diagnosing of candidal onychomycosis.

Publications

Translation elongation factor 1-alpha gene as a marker for diagnosing of candidal onychomycosis.

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