ArticleAntimicrobial agents and chemotherapy2012
Development of novel PCR assays to detect azole resistance-mediating mutations of the Aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients.
Article in Antimicrobial agents and chemotherapy, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. It is linked to 2 registered trials, which are not on this map. Cited by 30 papers, 1 of them a synthesis that pooled it.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Open, Prospective, Multicenter Trial to Evaluate the Clinical Significance of Combined Serological (Galactomannan ELISA, Beta D Glucan Assay) and Molecular (Nested Aspergillus PCR Assay, Real-time qPCR Assay, Multifungal DNA Microarray) Diagnostic Assays to Detect and Characterize Fungal Pathogens in Bronchoalveolar Lavage (BAL)and Blood Samples of Intensive Care Unit Patients With Pulmonary Infiltrates
Open, Prospective, Multicenter Trial to Evaluate the Clinical Significance of Combined Serological (Galactomannan ELISA, 1->3-β-D Glucan Assay) and Molecular (Nested Aspergillus PCR Assay, Multifungal DNA Microarray) Diagnostic Assays to Detect and Characterize Fungal Pathogens in Bronchoalveolar Lavage (BAL) and Blood Samples of Hematological High Risk Patients and to Detect Point Mutations Conferring Azole Resistance
Who cites it
30 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Aspergillus fumigatus-a systematic review to inform the World Health Organization priority list of fungal pathogens.Medical mycology · 2024Pooled it
- Molecular Mechanisms, Diagnosis, and Therapeutic Strategies of Antifungal Resistance in Filamentous Fungi.Journal of fungi (Basel, Switzerland) · 2026Review
- Azole Resistance inOpen forum infectious diseases · 2026Article
- Investigation of azole resistance inJournal of medical microbiology · 2026Article
- Amphotericin B in the Era of New Antifungals: Where Will It Stand?Journal of fungi (Basel, Switzerland) · 2024Review
- Citizen science reveals landscape-scale exposures to multiazole-resistantScience advances · 2023Article
- Detection of azole-resistant Aspergillus fumigatus in the environment from air, plant debris, compost, and soil.PloS one · 2023Article
- Species Distribution and Antifungal Susceptibilities ofJournal of clinical microbiology · 2022Article
- Review
- Molecular Markers of Antifungal Resistance: Potential Uses in Routine Practice and Future Perspectives.Journal of fungi (Basel, Switzerland) · 2021Review
- Identification of Fungal Species and Detection of Azole-Resistance Mutations in theYonsei medical journal · 2020Article
- Detecting Azole-Antifungal Resistance inJournal of fungi (Basel, Switzerland) · 2020Review
- (New) Methods for Detection ofCurrent fungal infection reports · 2019Article
- A simple and low cost tetra-primer ARMS-PCR method for detection triazole-resistant Aspergillus fumigatus.Molecular biology reports · 2019Article
- Azole Resistance in Aspergillus fumigatus in Patients with Cystic Fibrosis: A Matter of Concern?Mycopathologia · 2018Review
- Comparison of Two Molecular Assays for Detection and Characterization ofFrontiers in microbiology · 2018Article
- Molecular Detection of Azole-ResistantFrontiers in microbiology · 2018Review
- Performance of Molecular Approaches forFrontiers in microbiology · 2018Article
- Development and Validation of a High-Resolution Melting Assay To Detect Azole Resistance in Aspergillus fumigatus.Antimicrobial agents and chemotherapy · 2017Article
- Molecular Tools for the Detection and Deduction of Azole Antifungal Drug Resistance Phenotypes in Aspergillus Species.Clinical microbiology reviews · 2017Review
Corrections and comments
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The increasing incidence of azole resistance in Aspergillus fumigatus causing invasive aspergillosis (IA) in immunocompromised/hematological patients emphasizes the need to improve the detection of resistance-mediating cyp51A gene mutations from primary clinical samples, particularly as the diagnosis of invasive aspergillosis is rarely based on a positive culture yield in this group of patients. We generated primers from the unique sequence of the Aspergillus fumigatus cyp51A gene to establish PCR assays with consecutive DNA sequence analysis to detect and identify the A. fumigatus cyp51A tandem repeat (TR) mutation in the promoter region and the L98H and M220 alterations directly in clinical samples. After testing of the sensitivity and specificity of the assays using serially diluted A. fumigatus and human DNA, A. fumigatus cyp51A gene fragments of about 150 bp potentially carrying the mutations were amplified directly from primary clinical samples and subsequently DNA sequenced. The determined sensitivities of the PCR assays were 600 fg, 6 pg, and 4 pg of A. fumigatus DNA for the TR, L98H, and M220 mutations, respectively. There was no cross-reactivity with human genomic DNA detectable. Sequencing of the PCR amplicons for A. fumigatus wild-type DNA confirmed the cyp51A wild-type sequence, and PCR products from one azole-resistant A. fumigatus isolate showed the L98H and TR mutations. The second azole-resistant isolate revealed an M220T alteration. We consider our assay to be of high epidemiological and clinical relevance to detect azole resistance and to optimize antifungal therapy in patients with IA.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.