Rapid and Accurate Identification of &lt;b&gt;&lt;i&gt;Candida albicans&lt;/i&gt;&lt;/b&gt; and &lt;b&gt;&lt;i&gt;Candida dubliniensis&lt;/i&gt;&lt;/b&gt; by Real-Time PCR and Melting Curve Analysis

Asadzadeh, Mohammad; Ahmad, Suhail; Al‐Sweih, Noura; Khan, Ziauddin

doi:10.1159/000493426

Cited by 24 publications

(24 citation statements)

References 35 publications

(54 reference statements)

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“…Moreover, correct differentiation of this species is especially important for immunosuppressed patients, for example, HIV-infected patients, whose oral cavities are frequently colonized with C. dubliniensis [ 26 , 27 ]. In order to properly identify these species, it is necessary to introduce more advanced diagnostic methods that will standardize the tests using a more unified identification system, e.g., RT PCR [ 28 ].…”

Section: Discussionmentioning

confidence: 99%

Identification of Fungi Isolated from Oral Cavity of Patients with HIV Using MALDI–TOF MS

Pawlak

Andrusiów

Pajączkowska

et al. 2021

JCM

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Background: A growing incidence of invasive fungal infections, especially among immunocompromised patients, has given increased significance to microbiological diagnostics of yeast-like fungi. More accurate and faster fungi identification methods that can compete with classical methods are being searched for. In this paper, classical microbiological methods are compared to MALDI–TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry). Methods: The diagnostic material was collected from buccal mucosa from 98 adults, including 69 with HIV. Only positive cultures were included in the study. Results: Matching results were obtained in 45 samples, and there were nonmatching results in 35 samples, with the majority of these in the study group, constituting 50% of identifications within this group. A particularly common mistake resulting from the use of classical methods is the false identification of C. dubliniensis as C. albicans. Additionally, C. tropicalis proves to be difficult to identify. Conclusions: Our results and literature data suggest that MALDI–TOF MS should be considered an effective alternative to classical methods in terms of fungi identification, especially among HIV-positive patients, due to the different morphology of fungal colonies.

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Section: Discussionmentioning

confidence: 99%

Identification of Fungi Isolated from Oral Cavity of Patients with HIV Using MALDI–TOF MS

Pawlak

Andrusiów

Pajączkowska

et al. 2021

JCM

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“…In Somogyvari et al's study, 25 10 Candida spp were differentiated by analyzing the HRMA of the ITS2 sequence. In a study concluded by Asadzadeh et al, 26 real‐time PCR method based on melting point analysis with SYBR Green dye was introduced as a fast and reliable molecular method to perform accurate diagnosis and differentiation of C albicans and C dubliniensis . Alnuaimi et al 27 identified nine Candida spp using HRMA based on ITS1, 5.8S, and ITS2 sequences, in which they used eight Candida species as positive controls in each run.…”

Section: Discussionmentioning

confidence: 99%

Molecular identification of Candida isolates by Real‐time PCR‐high‐resolution melting analysis and investigation of the genetic diversity of Candida species

Nejad

Almani

Mohammadi

et al. 2020

Clinical Laboratory Analysis

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Background Candida species are considered as the cause of one of the most important opportunistic fungal diseases. Accurate identification of Candida species is important because of antifungal susceptibility patterns are different among these species, so proper identification helps in the selection of antifungal drugs for the prevention and treatment. Phenotypic methods for identification of Candida species, which are widely used in clinical microbiology laboratories, have some limitations. Real‐time PCR followed by the high‐resolution melting analysis (HRMA) is a novel approach for the rapid recognition of pathogenic fungi. Molecular phylogeny is essential for obtaining a better understanding of the evolution of the genus Candida and the identification of the relative degree of the Candida species. The purpose of this study was molecular identification of Candida isolates by Real‐time PCR‐high‐resolution melting analysis and investigation of the genetic diversity of Candida species. Methods Two hundred and thirty‐two Candida isolates including 111 Candida isolates obtained from 96 HIV/AIDS patients and 121 Candida isolates obtained from 98 non‐HIV persons were identified by real‐time PCR and high‐resolution melting curve analysis. To evaluate genetic diversity and relationships among Candida species, PCR products of nine clinical Candida isolates, as a representative of each kind of species, were randomly selected for DNA sequence analysis. Results In HIV/AIDS patients, six species of Candida spp. were identified as follows: C albicans (n = 64; 57.7%), C glabrata (n = 31; 27.92%), C parapsilosis (n = 9; 8.1%), C tropicalis (n = 4; 3.6%), C krusei (n = 2; 1.8%), and C kefyr (n = 1; 0.90%). In non‐HIV persons, we identified eight species of Candida including C albicans (n = 46; 38.33%) followed by C glabrata and C krusei (each one, n = 18; 15%), C tropicalis (n = 13; 10.83%), C lusitaniae (n = 12; 5.17%), C parapsilosis (n = 10; 4.31%), and C kefyr and C guillermondii (each one, n = 2; 1.66%). Also, the phylogenetic analysis showed the presence of two main clades and six separate subclades. Accordingly, about 88.9% of the isolates were located in clade I and 11.10% of the studied isolates were in clade II. Conclusions Real‐time PCR followed by high‐resolution melting analysis (HRMA) is known as a reliable, fast, and simple approach for detection and accurate identification of Candida species, especially in clinical samples.

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“…and C. albicans genotypes [230]. Furthermore, Asadzadeh et al [231] developed a duplex RT-PCR assay for rapid detection and differentiation between C. albicans and C. dubliniensis by using two species-specific primer pairs and SYBR ® Green dye to differentiate C. albicans and C. dubliniensis isolates via melting curve analysis of RT-PCR amplicons. The amplification products were also analyzed by agarose gel electrophoresis to confirm RT-PCR results.…”

Section: Molecular Methods Used To Detect Yeastsmentioning

confidence: 99%

“…The amplification products were also analyzed by agarose gel electrophoresis to confirm RT-PCR results. Melting temperature (Tm) for reference strains of C. albicans and C. dubliniensis were 86.55 °C and 82.75 °C, respectively [231]. Similarly, quantitative PCR assays to determine the relative Paracoccidioides brasiliensis load in lungs from infected mice were also developed.…”

Section: Molecular Methods Used To Detect Yeastsmentioning

confidence: 99%

Advances in Chemical and Biological Methods to Identify Microorganisms—From Past to Present

et al. 2019

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Fast detection and identification of microorganisms is a challenging and significant feature from industry to medicine. Standard approaches are known to be very time-consuming and labor-intensive (e.g., culture media and biochemical tests). Conversely, screening techniques demand a quick and low-cost grouping of bacterial/fungal isolates and current analysis call for broad reports of microorganisms, involving the application of molecular techniques (e.g., 16S ribosomal RNA gene sequencing based on polymerase chain reaction). The goal of this review is to present the past and the present methods of detection and identification of microorganisms, and to discuss their advantages and their limitations.

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Rapid and Accurate Identification of <b><i>Candida albicans</i></b> and <b><i>Candida dubliniensis</i></b> by Real-Time PCR and Melting Curve Analysis

Cited by 24 publications

References 35 publications

Identification of Fungi Isolated from Oral Cavity of Patients with HIV Using MALDI–TOF MS

Identification of Fungi Isolated from Oral Cavity of Patients with HIV Using MALDI–TOF MS

Molecular identification of Candida isolates by Real‐time PCR‐high‐resolution melting analysis and investigation of the genetic diversity of Candida species

Advances in Chemical and Biological Methods to Identify Microorganisms—From Past to Present

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