ArticleJournal of clinical microbiology2008
Automated and manual methods of DNA extraction for Aspergillus fumigatus and Rhizopus oryzae analyzed by quantitative real-time PCR.
Article in Journal of clinical microbiology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- Amplification of RNA for identification of Zika and HCV in whole blood.Science advances · 2026Article
- Enhancing Clinical Utility: Utilization of International Standards and Guidelines for Metagenomic Sequencing in Infectious Disease Diagnosis.International journal of molecular sciences · 2024Review
- Establishment and Characterization of the First Chinese National Standard for Nucleic Acid Amplification Technology Assays for Hepatitis E Virus Nucleic Acid Detection.Pathogens (Basel, Switzerland) · 2023Article
- Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection.Sensors (Basel, Switzerland) · 2021Article
- Establishment of a New PNA-FISH Method forMicroorganisms · 2020Article
- Fatal Pulmonary and Cerebellar Zygomycosis due to Rhizomucor pusillus in a Ringed Seal (Pusa hispida).Mycopathologia · 2018Article
- Evaluation of Two Commercial Real-Time PCR Kits for Aspergillus DNA Detection in Bronchoalveolar Lavage Fluid in Patients with Invasive Pulmonary Aspergillosis.The Journal of molecular diagnostics : JMD · 2018Article
- Article
- Identification of fungal pathogens in a patient with acute myelogenic leukemia using a pathogen detection array technology.Cancer biology & therapy · 2016Article
- Effort versus Reward: Preparing Samples for Fungal Community Characterization in High-Throughput Sequencing Surveys of Soils.PloS one · 2015Article
- Rapid detection and identification of mucormycetes in bronchoalveolar lavage samples from immunocompromised patients with pulmonary infiltrates by use of high-resolution melt analysis.Journal of clinical microbiology · 2014Article
- Molecular and nonmolecular diagnostic methods for invasive fungal infections.Clinical microbiology reviews · 2014Review
- Streamlining DNA barcoding protocols: automated DNA extraction and a new cox1 primer in arachnid systematics.PloS one · 2014Article
- Locked-in syndrome after basilary artery thrombosis by mucormycosis masquerading as meningoencephalitis in a lymphoma patient.Ecancermedicalscience · 2013Article
- Optimization of Toxoplasma gondii DNA extraction from amniotic fluid using NucliSENS easyMAG and comparison with QIAamp DNA minikit.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2012Article
- Simultaneous detection and identification of Aspergillus and mucorales species in tissues collected from patients with fungal rhinosinusitis.Journal of clinical microbiology · 2011Article
- Recent advances in the treatment of mucormycosis.Current infectious disease reports · 2010Article
- Detection of Aspergillus fumigatus in a rat model of invasive pulmonary aspergillosis by real-time nucleic acid sequence-based amplification.Journal of clinical microbiology · 2010Article
- Recent advances in the management of mucormycosis: from bench to bedside.Clinical infectious diseases : an official publication of the Infectious Diseases Society of America · 2009Review
- Diagnosis of invasive aspergillosis using bronchoalveolar lavage in haematology patients: influence of bronchoalveolar lavage human DNA content on real-time PCR performance.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2009Article
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8 authors.
Funding
Abstract
Quantitative real-time PCR (qPCR) may improve the detection of fungal pathogens. Extraction of DNA from fungal pathogens is fundamental to optimization of qPCR; however, the loss of fungal DNA during the extraction process is a major limitation to molecular diagnostic tools for pathogenic fungi. We therefore studied representative automated and manual extraction methods for Aspergillus fumigatus and Rhizopus oryzae. Both were analyzed by qPCR for their ability to extract DNA from propagules and germinated hyphal elements (GHE). The limit of detection of A. fumigatus and R. oryzae GHE in bronchoalveolar lavage (BAL) fluid with either extraction method was 1 GHE/ml. Both methods efficiently extracted DNA from A. fumigatus, with a limit of detection of 1 x 10(2) conidia. Extraction of R. oryzae by the manual method resulted in a limit of detection of 1 x 10(3) sporangiospores. However, extraction with the automated method resulted in a limit of detection of 1 x 10(1) sporangiospores. The amount of time to process 24 samples by the automated method was 2.5 h prior to transferring for automation, 1.3 h of automation, and 10 min postautomation, resulting in a total time of 4 h. The total time required for the manual method was 5.25 h. The automated and manual methods were similar in sensitivity for DNA extraction from A. fumigatus conidia and GHE. For R. oryzae, the automated method was more sensitive for DNA extraction of sporangiospores, while the manual method was more sensitive for GHE in BAL fluid.
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