Application of High-Resolution Melting PCR to Detect the Genomic Fungal ITS 2 Region

Nawar, Nada; Behiry, Iman Kamal; Yousef, Reham H. A.; Emara, Mohamed

doi:10.1093/labmed/lmz034

Cited by 4 publications

(2 citation statements)

References 25 publications

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“…Similar to our results, Lau et al (17) showed that the RT-PCR had a higher sensitivity for detecting IFIs than blood culture tests. In contrast, Nawar et al (18) reported that HRM-PCR had a lower sensitivity for detecting IFIs than the blood culture technique.…”

Section: Discussionmentioning

confidence: 94%

Evaluation of Panfungal Polymerase Chain Reaction in Early Detection of Invasive Fungal Infections in Egyptian Patients with Hematological Malignancies

et al. 2022

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The incidence of invasive fungal infections (IFIs) has increased owing to the rising number of immunodeficient patients. A case-control study was performed at the Ain Shams University Hospitals, Cairo, Egypt. The case group (n = 80) included 80 patients diagnosed with hematological malignancies, and the control group (n = 20) included 20 patients. All patients were tested for the presence of fungal species using blood culture and panfungal real-time polymerase chain reaction (RT-PCR). Fungal species differentiation was performed using high-resolution melting (HRM) PCR. There were 39 suspected cases of IFIs among the 80 patients. The panfungal RT-PCR detection rate was 51.3% (41/80). HRM-PCR identified that 51.2% of the fungal species were Candida albicans, 44.0% were non-Candida albicans, and 4.9% were Mucor. The blood cultures were positive for the presence of fungi in two patients with acute myeloid leukemia. The fungal detection rate using the panfungal RT-PCR technique was significantly higher than that using the blood culture technique (P < 0.001). RT-PCR using panfungal markers is sensitive, rapid, and superior to the blood culture technique to detect IFIs. HRM-PCR is a specific test for species identification.

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Section: Discussionmentioning

confidence: 94%

Evaluation of Panfungal Polymerase Chain Reaction in Early Detection of Invasive Fungal Infections in Egyptian Patients with Hematological Malignancies

et al. 2022

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“…Günümüzde bazı araştırmacılar Candida türlerinin belirlenmesinde RT-PCR HRMA yöntemini tercih etmektedir (32,34,38,39) . Nemcova ve ark.…”

Section: Candida Parapsilosis -13 13unclassified

Diagnostic Value of The Real-Time PCR Test for The Identification of Candida Species in Oral Rinse Solutions

Çürük,

Karakoyun,

Ünal

et al. 2023

Turk Mikrobiyol Cemiy Derg

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Objective: A variety of microorganisms are found in the oral cavities of healthy humans, including yeast fungi. Disruption of the balance among these microorganisms may cause excessive yeast colonization, or oral candidosis. In the present study, we aimed to investigate the diagnostic value of a real-time PCR method for the identification of medically important Candida species in the oral flora. Methods: Two hundred students from Çukurova University Faculty of Dentistry participated in this study. The concentrated oral rinse water samples from the participants were inoculated into CHROMagar Candida (CAC) medium with a swab. CAC along with RT-PCR and MALDI-TOF MS were used to identify the Candida species. Results: A total of 68 yeast fungi were isolated from 52 (26%) clinical samples. The most common isolated species were C. albicans (36.1%), C. parapsilosis (21.3%), C. dubliniensis (14.7%), C. lusitaniae (9.8%), C. kefyr (8.2%), C. lambica (3.3%) and other Candida species (6.6%). Oral colonisation of yeast fungi was associated with the toothpaste brand (p=0.02). The Candida species isolated in this study were reliably identified by RT-PCR melting curve analysis (MCA), except C. albicans, C. kefyr and C. lambica. However, this method could not distinguish C. albicans and C. kefyr based on their peak melting temperatures (Tm). Conclusion: It may take 2-3 days for Candida species to be identified and differentiated from each other using conventional methods. MCA of Candida species using RT-PCR, which can be accomplished in a single day, is a simple, inexpensive, and rapid method. However, this study revealed that RT-PCR was not reliable in the identification of Candida species (C. albicans, C. kefyr, and C. lambica).

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Molecular Diagnosis of Yeast Infections

White

Price

Cordey

et al. 2021

Curr Fungal Infect Rep

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Purpose of Review The use of molecular tests to aid the diagnosis of invasive yeast infection, in particular invasive candidosis, has been described for over two decades, yet widespread application is limited, and diagnosis remains heavily dependent on classical microbiology. This article will review developments from the past decade in attempt to build on existing knowledge. It will highlight clinical performance and limitations while reviewing developments on recognized procedures; it will also provide insight into novel approaches incorporated in response to clinical demand (e.g. C. auris and antifungal resistance) or technological advances (e.g. next-generation sequencing). Recent Findings Limited methodological standardization and, until recently, unavailability of commercial options have hindered the integration of molecular diagnostics for yeasts. The development of certain, novel commercial methods has received considerable evaluation allowing a greater understanding of individual assay performance, but widespread multicentre evaluation of most commercial kits is lacking. The detection of emerging pathogens (e.g. C. auris ) has been enhanced by the development of molecular tests. Molecular methods are providing a better understanding of the mycobiome, mechanisms of resistance and epidemiology/phylogeny. Summary Despite over two decades of use, the incorporation of molecular methods to enhance the diagnosis of yeast infections remains limited to certain specialist centres. While the development of commercial tests will provide stimulus for broader application, further validation and reduced costs are required. Over the same period of time, Aspergillus PCR has become more widely accepted driven by international efforts to standardize methodology; it is critical that yeast PCR follows suit. Next-generation sequencing will provide significant information on the mycobiome, antifungal resistance mechanism and even broad-range detection directly from the specimen, which may be critical for the molecular detection of yeasts other than Candida species, which is currently limited.

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Application of High-Resolution Melting PCR to Detect the Genomic Fungal ITS 2 Region

Cited by 4 publications

References 25 publications

Evaluation of Panfungal Polymerase Chain Reaction in Early Detection of Invasive Fungal Infections in Egyptian Patients with Hematological Malignancies

Evaluation of Panfungal Polymerase Chain Reaction in Early Detection of Invasive Fungal Infections in Egyptian Patients with Hematological Malignancies

Diagnostic Value of The Real-Time PCR Test for The Identification of Candida Species in Oral Rinse Solutions

Molecular Diagnosis of Yeast Infections

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