Different Methods Compared for Isolating Fungal DNA
|
By LabMedica International staff writers Posted on 26 Apr 2021 |
|
Image: A 47-year-old man with mucormycosis and (b) electron micrograph of his skin showing sporangia of Mucorales fungi: non-apophysate sporangia with pronounced columellae and conspicuous collarette at the base of the columella following sporangiospore dispersal (Photo courtesy of IntechOpen Limited)
Mucormycosis (previously called zygomycosis) is a serious, but rare fungal infection, caused by a group of molds called mucormycetes. These molds live throughout the environment. Mucormycosis mainly affects people who are immunocompromised or diabetic.
Mucormycosis frequently infects the sinuses, brain, or lungs. While infection of the oral cavity or brain are the most common forms of mucormycosis, the fungus can also infect other areas of the body such as the gastrointestinal tract, skin, and other organ systems. The disease is often characterized by hyphae growing in and around blood vessels.
Medical Scientists at the University of Turin School of Medicine (Turin, Italy) compare the methods of DNA isolation in the moulds of genus Mucorales with special regard to the amount and purity of the DNA acquired. A total of six Mucorales clinical strains were used and these included three Mucor spp. and three Rhizopus spp. Filamentous fungi were identified according to macroscopic and microscopic morphological procedures and maintained on potato dextrose agar. For DNA extraction five named methods A, B, C, D and E protocol were compared. Amplification and amplicon detection using extracted DNA template was performed using an Applied Biosystems 7500 Real Time PCR System (Life Technologies, Carlsbad, CA, USA).
The scientists reported that method A based on boiling moulds obtained a greater amount of fungal DNA, but failed to extract amplifiable DNA from Mucorales. Microwave and freezing, peculiarity of methods B and C respectively, have achieved poor results both in terms of DNA concentration and of amplification. On the other hand, method E based on addition of a potassium acetate buffer in the lysis step, obtained a decent level of amount of fungal DNA with greater amplifiable DNA from Mucorales.
The authors concluded that overall, the E method was the most efficient method in the extraction of DNA from fungal cultures compared to the other methods considering time, cost, technical expertise, and instrumentation. Use of this assay will allow medical laboratorians to obtain DNA from fungi quickly for use in molecular assays. The study was published on April 17, 2021 in the journal Practical Laboratory Medicine.
Related Links:
University of Turin School of Medicine
Life Technologies
Mucormycosis frequently infects the sinuses, brain, or lungs. While infection of the oral cavity or brain are the most common forms of mucormycosis, the fungus can also infect other areas of the body such as the gastrointestinal tract, skin, and other organ systems. The disease is often characterized by hyphae growing in and around blood vessels.
Medical Scientists at the University of Turin School of Medicine (Turin, Italy) compare the methods of DNA isolation in the moulds of genus Mucorales with special regard to the amount and purity of the DNA acquired. A total of six Mucorales clinical strains were used and these included three Mucor spp. and three Rhizopus spp. Filamentous fungi were identified according to macroscopic and microscopic morphological procedures and maintained on potato dextrose agar. For DNA extraction five named methods A, B, C, D and E protocol were compared. Amplification and amplicon detection using extracted DNA template was performed using an Applied Biosystems 7500 Real Time PCR System (Life Technologies, Carlsbad, CA, USA).
The scientists reported that method A based on boiling moulds obtained a greater amount of fungal DNA, but failed to extract amplifiable DNA from Mucorales. Microwave and freezing, peculiarity of methods B and C respectively, have achieved poor results both in terms of DNA concentration and of amplification. On the other hand, method E based on addition of a potassium acetate buffer in the lysis step, obtained a decent level of amount of fungal DNA with greater amplifiable DNA from Mucorales.
The authors concluded that overall, the E method was the most efficient method in the extraction of DNA from fungal cultures compared to the other methods considering time, cost, technical expertise, and instrumentation. Use of this assay will allow medical laboratorians to obtain DNA from fungi quickly for use in molecular assays. The study was published on April 17, 2021 in the journal Practical Laboratory Medicine.
Related Links:
University of Turin School of Medicine
Life Technologies
Platinum Member
Automated Coagulation Analyzer
Hemolumi H6
New
Neurofilament Light Chain Assay
Lumipulse G NfL Blood
Gold Member
Electrolyte Analyzer
CBS-4000 (CBS-400)
Latest Microbiology News
- Molecular Panel Gains WHO Emergency Use Listing to Support Ebola Outbreak Response
- Rapid Host-Response Test Aims to Predict Corticosteroid Benefit in Sepsis
- Oral and Lung Microbiome Changes Linked to Severe Long Covid Symptoms
- Study Finds Distinct Malaria Parasites Linked to Sickle Cell Trait Protection
- Rapid Urine Test Aids Diagnosis of Invasive Aspergillosis
- Nanodroplet CRISPR Technology Supports Rapid, Multiplexed Mycobacterial Identification
- Genomic Workflow Identifies Fungal Pathogens Before Blood Cultures Turn Positive
- FDA-Cleared Multiplex PCR Test Detects 13 Respiratory Pathogens in a Single Sample
- One-Hour Molecular Panel Expands Bloodstream Infection Testing for Gram-Negative Pathogens
- FDA Clears Rapid Phenotypic Antimicrobial Susceptibility System for Positive Blood Cultures
- New Urine Test Expands Mycotoxin Analysis to 31 Markers for Broader Exposure Assessment
- Collaboration Targets Blood-Based Ebola Detection Outside Central Laboratories
- Study Finds Frequent Overlap in Antibodies to Lyme and Other Tick-Borne Pathogens
- Early-Life Gut Microbiome Changes May Help Assess Type 1 Diabetes Risk
- FDA Breakthrough-Designated Urine Panel Aims to Shorten Pathogen and Susceptibility Testing
- Expanded Newborn Screening Program Increases Detection of Congenital Cytomegalovirus
Channels
Clinical Chemistry
view channel
Daily Blood Biomarker Testing May Help Guide Earlier Antibiotic Stopping in Sepsis
Sepsis is a life-threatening condition caused by the body’s overwhelming response to infection and accounts for thousands of deaths each year. In intensive care, concern about stopping treatment too early... Read more
Blood Proteins Reveal Signals Linked to Prostate Cancer Risk Years Before Diagnosis
Prostate cancer is the most commonly diagnosed cancer in men worldwide. While many tumors grow slowly, aggressive forms can be life-threatening, underscoring the need for earlier risk identification.... Read more
Two-Step Blood Testing Approach Reduces Uncertainty in Amyloid Assessment
Blood-based biomarkers offer a more accessible, less expensive way to detect Alzheimer’s-related brain changes than traditional methods such as amyloid-beta positron emission tomography (Aβ-PET).... Read more
Multimodal Platform to Combine Blood and Imaging Biomarkers for Lung Nodule Assessment
Lung nodules are commonly detected during chest imaging and require careful risk assessment to determine whether follow-up imaging, additional testing, or biopsy is needed. With millions of patients evaluated... Read moreMolecular Diagnostics
view channel
AI Tool Predicts Immunotherapy Response Using Tumor Senescence Signals
Patients receiving the same cancer immunotherapy can have markedly different outcomes, with some showing little or no response. Identifying these patients earlier could reduce the costs of ineffective... Read more
New Variant Tracking Option Expands Molecular Residual Disease Monitoring
Molecular residual disease (MRD) testing can detect traces of cancer that persist after treatment or emerge during recurrence. Because circulating tumor DNA (ctDNA) may be present at very low levels, residual... Read more
Hematology
view channel
New Myelofibrosis Guidelines Recommend Expanded Genetic Testing at Diagnosis
Myelofibrosis is a rare, life-threatening blood cancer affecting up to six per 100,000 people. The disease causes scarring in the bone marrow, limiting the production of healthy blood cells and potentially... Read more
Handheld Point-of-Care Test Aims to Enable Rapid Coagulation Assessment in Trauma
Severe blood loss and inadequate tissue oxygenation are leading causes of preventable death after traumatic injury. Blood products are essential in trauma care, but cold-chain storage, short shelf life,... Read more
Immunology
view channel
Antibody Profiling Predicts Inflammatory Bowel Disease Years Before Diagnosis
Inflammatory bowel disease (IBD), which includes Crohn’s disease and ulcerative colitis, involves abnormal immune responses that cause persistent inflammation and damage in the digestive system.... Read more
Immune Profiling Initiative Aims to Improve Bone Marrow Analysis in Leukemia
Acute myeloid leukemia (AML) is an aggressive blood cancer in which assessing disease burden after therapy can help guide clinical evaluation. Patients with measurable residual disease (MRD) after treatment... Read more
Immune Profiling Identifies Early Maternal Autoantibodies Linked to Preterm Birth Risk
A typical pregnancy lasts about 40 weeks, but labor begins prematurely in roughly one in 10 pregnancies. Preterm birth is the leading cause of newborn death worldwide, yet its underlying causes and which... Read more
Longitudinal Blood Testing Reveals Changes in Allergen Sensitization During Childhood
Food allergy is a major childhood health concern, and distinguishing persistent risk from transient sensitization can guide clinical decisions. Specific IgE antibodies may be detectable even when a child... Read morePathology
view channel
AI Tissue Analysis May Help Identify Missed Precancerous Colon Polyps
Colorectal cancer is diagnosed in an estimated 1.9 million people worldwide each year, often without symptoms in its early stages. Timely detection of precancerous lesions is therefore important, yet some... Read more
AI Pre-Screening Helps Identify Lung Cancer Cases for Additional Biomarker Testing
Tempus AI (Chicago, IL, USA) will present findings from a study evaluating an artificial intelligence (AI)-enabled pre-screening panel for advanced non-small cell lung cancer (NSCLC) at the ESMO Congress... Read more
AI Pathology Platform Targets Workflow Delays and Molecular Test Failures
Pathology laboratories often rely on visual review of tissue slides to assess quality, identify tumors, count mitoses, and estimate tumor content. These tasks can be time-consuming and vary between readers,... Read more
Molecular Testing Refines Barrett’s Esophagus Progression Risk Beyond Biopsy Findings
Barrett’s esophagus is a precancerous condition associated with increased risk of esophageal adenocarcinoma. Routine endoscopic biopsy findings guide surveillance and therapy, but histopathology may not... Read moreTechnology
view channel
New AI-Powered System Aims to Streamline Laboratory and Manufacturing Workflows
Scientific laboratories supporting drug development, biotechnology, and materials research increasingly rely on coordinated digital workflows to move data, samples, and activities across teams.... Read more
Laser-Based Swab Analysis Shows Promise for Detecting Disease-Linked Odor Patterns
Disease-related changes in volatile organic compounds can alter body odor, producing measurable patterns in exhaled breath and bodily fluids. Current analytical methods can be complex, time-consuming,... Read more
Industry
view channel
Collaboration to Expand Cervical Cancer Screening Access With At-Home HPV Collection
Cervical cancer is largely preventable with regular screening and early detection of human papillomavirus (HPV), the leading cause of the disease. Screening gaps persist, in part because traditional sample... Read more
Alzheimer’s Progression Blood Test Gains U.S. Patent Ahead of Planned Launch
Alzheimer’s disease is the most common cause of dementia, accounting for an estimated 60–70% of cases worldwide. Its progressive effects on memory, thinking, behavior, and daily functioning underscore... Read more
