ArticlePloS one2013
Comparison of pathogen DNA isolation methods from large volumes of whole blood to improve molecular diagnosis of bloodstream infections.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 42 papers, 1 of them a synthesis that pooled it.
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Who cites it
42 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Pooled it
- Diagnostic performance of a loop-mediated isothermal amplification test for the diagnosis of human and cat sporotrichosis in two highly endemic areas in Brazil: insights from a real-world, field-based evaluation.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2026Article
- Fast Microbiology: A Pilot Study Comparing Culture-Independent Digital PCR and Blood Culture for Pathogen Detection in Sepsis.International journal of molecular sciences · 2026Article
- Comparison evaluation of bacterial DNA extraction methods for improved molecular diagnostic accuracy of sepsis-causing pathogens in clinical whole blood samples.Scientific reports · 2025Article
- Optimizing sample preparation for culture-free nanopore sequencing to enable rapid pathogen and antimicrobial resistance profiling in bovine mastitis.Frontiers in microbiology · 2025Article
- Randomized Trial Demonstrating No Translocation of Intact Intestinal Bacteria During Hemodialysis or Hemodiafiltration.Kidney international reports · 2025Article
- Assessing Diagnostic Performance of Molecular Culture for Neonatal Sepsis: Protocol of the CHAMPIONS Study.Diagnostics (Basel, Switzerland) · 2024Article
- Development of two multiplex PCR assays for rapid detection of eleven Gram-negative bacteria in children with septicemia.Tropical medicine and health · 2024Article
- Potential of Molecular Culture in Early Onset Neonatal Sepsis Diagnosis: A Proof of Principle Study.Microorganisms · 2023Article
- Establishment of a Fast Diagnostic Method for Sepsis Pathogens Based on M1 Bead Enrichment.Current microbiology · 2023Article
- Review
- Multiplex Microsphere PCR (mmPCR) Allows Simultaneous Gram Typing, Detection of Fungal DNA, and Antibiotic Resistance Genes.Laboratory medicine · 2022Article
- Bacterial DNA load in Staphylococcus aureus bacteremia is significantly higher in intravascular infections.PloS one · 2022Article
- Direct detection of microorganisms in sonicated orthopedic devices after in vitro biofilm production and different processing conditions.European journal of orthopaedic surgery & traumatology : orthopedie traumatologie · 2021Article
- Optimized bacterial DNA isolation method for microbiome analysis of human tissues.MicrobiologyOpen · 2021Article
- Molecular Diagnosis of Yeast Infections.Current fungal infection reports · 2021Review
- DNA/RNA Electrochemical Biosensing Devices a Future Replacement of PCR Methods for a Fast Epidemic Containment.Sensors (Basel, Switzerland) · 2020Review
- PCR-detectableOpen life sciences · 2020Article
- Diagnostic Performance of a Novel Multiplex PCR Assay for Candidemia among ICU Patients.Journal of fungi (Basel, Switzerland) · 2019Article
- Neutrophil-to-Lymphocyte and Platelet-to-Lymphocyte Ratios Are Correlated with Complicated Diverticulitis and Hinchey Classification: A Simple Tool to Assess Disease Severity in the Emergency Department.Emergency medicine international · 2019Article
Corrections and comments
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Authors and funding
11 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
For patients suffering from bloodstream infections (BSI) molecular diagnostics from whole blood holds promise to provide fast and adequate treatment. However, this approach is hampered by the need of large blood volumes. Three methods for pathogen DNA isolation from whole blood were compared, i.e. an enzymatic method (MolYsis, 1-5 ml), the novel non-enzymatic procedure (Polaris, 1-5 ml), and a method that does not entail removal of human DNA (Triton-Tris-EDTA EasyMAG, 200 µl). These methods were evaluated by processing blood spiked with 0-1000 CFU/ml of Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans. Downstream detection was performed with real-time PCR assays. Polaris and MolYsis processing followed by real-time PCRs enabled pathogen detection at clinically relevant concentrations of 1-10 CFU/ml blood. By increasing sample volumes, concurrent lower cycle threshold (Ct) values were obtained at clinically relevant pathogen concentrations, demonstrating the benefit of using larger blood volumes. A 100% detection rate at a concentration of 10 CFU/ml for all tested pathogens was obtained with the Polaris enrichment, whereas comparatively lower detection rates were measured for MolYsis (50-67%) and EasyMAG (58-79%). For the samples with a concentration of 1 CFU/ml Polaris resulted in most optimal detection rates of 70-75% (MolYsis 17-50% and TTE-EasyMAG 20-36%). The Polaris method was more reproducible, less labour intensive, and faster (45 minutes (including Qiagen DNA extraction) vs. 2 hours (MolYsis)). In conclusion, Polaris and MolYsis enrichment followed by DNA isolation and real-time PCR enables reliable and sensitive detection of bacteria and fungi from 5 ml blood. With Polaris results are available within 3 hours, showing potential for improved BSI diagnostics.
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