Related Experiment Video
Updated: Apr 9, 2026
Whole Genome Sequencing of Candida glabrata for Detection of Markers of Antifungal Drug Resistance
Published on: December 28, 2017
Clinical impact of microbial cell-free DNA next-generation sequencing for invasive mold infection-a single-center
Rebecca Berger1, Nina Howard2, Sarah Grant2
1School of Medicine, Baylor College of Medicine, 1 Baylor Plaza, Houston, TX 77030, USA.
Background:
Diagnosing invasive mold infections (IMI) is challenging because they typically occur as opportunistic infections in immunocompromised patients who often present with non-specific symptoms. Furthermore, no single test can definitively diagnose IMI, and a proven diagnosis often requires invasive sampling. This sampling can be unsafe and difficult to perform, especially in the immunocompromised population most at risk for these infections.
Objectives:
The objective of this study is to assess the clinical utility of plasma microbial cell-free DNA next-generation sequencing (mcfDNA-NGS) for diagnosing invasive mold infections in the context of conventional diagnostic methods.
Design:
Retrospective observational study at a quaternary care center (2017-2025).
Methods:
The charts of 30 patients with mold-positive mcfDNA-NGS (Karius™ Spectrum; Redwood City, CA, USA) were reviewed, with IMI adjudicated per 2020 EORTC/MSGERC criteria. Provider documentation, medication orders, and patient outcomes were used to assess clinician interpretation of mcfDNA-NGS result accuracy (true positive vs false positive) and its impact on diagnostic and therapeutic decision-making. Turnaround time (TAT) and molecules per microliter (MPM) were summarized.
Results:
IMI final classifications were proven (6), probable (8), possible (5), and unclassified (11). Overall, 23/30 (77%) results were true positives. Among these 23 patients with true positive results for mold, 5 (22%) received a new clinical adjudication of IMI that had been entirely missed by conventional diagnostic testing. Furthermore, mcfDNA NGS provided species-level pathogen identification in 9 of the 23 (39%) true positive cases where conventional testing detected fungal elements or elevated biomarkers but could not identify the specific organism. Median TAT was 102 h; median MPM 657. MPM did not differ between true and false positives (p = 0.86). mcfDNA-NGS changed diagnostic classification in 7/30 (23%) and antimicrobial management in 16/30 (53%).
Conclusion:
mcfDNA-NGS provided noninvasive, actionable information, informing diagnosis and therapy. Future studies should define optimal stewardship and cost-effectiveness.
Insights
Plasma microbial cell-free DNA next-generation sequencing (mcfDNA-NGS) aids in diagnosing invasive mold infections (IMI). This noninvasive test accurately identified IMI missed by conventional methods, improving patient management.
Area of Science:
- Medical Diagnostics
- Infectious Diseases
- Molecular Biology
Background:
- Invasive mold infections (IMI) are challenging to diagnose in immunocompromised patients due to non-specific symptoms.
- Conventional diagnostics often require invasive sampling, posing risks.
- No single test definitively diagnoses IMI.
Purpose of the Study:
- To evaluate the clinical utility of plasma microbial cell-free DNA next-generation sequencing (mcfDNA-NGS) for diagnosing IMI.
- To compare mcfDNA-NGS with conventional diagnostic methods.
Main Methods:
- Retrospective observational study of 30 patients with mold-positive mcfDNA-NGS results.
- Adjudication of IMI based on 2020 EORTC/MSGERC criteria.
- Assessment of mcfDNA-NGS accuracy, impact on decision-making, turnaround time (TAT), and molecules per microliter (MPM).
Main Results:
- 77% of mcfDNA-NGS results were true positives.
- 5% of true positive cases had IMI missed by conventional diagnostics.
- mcfDNA-NGS provided species-level identification in 39% of true positive cases.
- Diagnostic classification changed in 23% and antimicrobial management in 53% of patients.
Conclusions:
- mcfDNA-NGS offers noninvasive, actionable data for IMI diagnosis and therapy.
- Further research is needed to establish optimal stewardship and cost-effectiveness.
