Metagenomic identification of severe pneumonia pathogens in mechanically-ventilated patients: a feasibility and clinical validity study
Year: 2019
Type: article
Study design: Observational Study
Abstract: BACKGROUND: Metagenomic sequencing of respiratory microbial communities for pathogen identification in pneumonia may help overcome the limitations of culture-based methods. We examined the feasibility and clinical validity of rapid-turnaround metagenomics with Nanopore™ sequencing of clinical respiratory specimens. METHODS: We conducted a case-control study of mechanically-ventilated patients with pneumonia (nine culture-positive and five culture-negative) and without pneumonia (eight controls). We collected endotracheal aspirates and applied a microbial DNA enrichment method prior to metagenomic sequencing with the Oxford Nanopore MinION device. For reference, we compared Nanopore results against clinical microbiologic cultures and bacterial 16S rRNA gene sequencing. RESULTS: Human DNA depletion enabled in depth sequencing of microbial communities. In culture-positive cases, Nanopore revealed communities with high abundance of the bacterial or fungal species isolated by cultures. In four cases with resistant clinical isolates, Nanopore detected antibiotic resistance genes corresponding to the phenotypic resistance in antibiograms. In culture-negative pneumonia, Nanopore revealed probable bacterial pathogens in 1/5 cases and Candida colonization in 3/5 cases. In controls, Nanopore showed high abundance of oral bacteria in 5/8 subjects, and identified colonizing respiratory pathogens in other subjects. Nanopore and 16S sequencing showed excellent concordance for the most abundant bacterial taxa. CONCLUSIONS: We demonstrated technical feasibility and proof-of-concept clinical validity of Nanopore metagenomics for severe pneumonia diagnosis, with striking concordance with positive microbiologic cultures, and clinically actionable information obtained from sequencing in culture-negative samples. Prospective studies with real-time metagenomics are warranted to examine the impact on antimicrobial decision-making and clinical outcomes.
Source: Respiratory Research
Language: English
Subfield: Clinical Biochemistry
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,570
Funders: Foundation for the National Institutes of Health, University of Pittsburgh, Oxford Nanopore Technologies, National Institute for Health and Care Research, Rosetrees Trust, National Institutes of Health, Directorate for Biological Sciences, Medical Center, University of Pittsburgh, Medical Research Council, Biotechnology and Biological Sciences Research Council, National Heart, Lung, and Blood Institute, National Institute of General Medical Sciences
Awards: G2210635924, Pathogenesis of Obstruction/Emphysema and the Microbiome (POEM) in HIV, Epidemiology and Evolution of Pathogens in the Food Chain, G2506595212, G2561744426, G3917667430, G4245513023, INHALE: Potential of Molecular Diagnostics for Hospital-Acquired and Ventilator-Associated Pneumonia in UK Critical Care, G5351278108, G5534850370, G5728561071, G6138878392, Microbial Survival in the Food Chain, G6358829866, G7572515059, G8430620755, G8814408722, G8926217196, G9000002626, G9000018726, G9000030573, G9000122246, G9000189855, Sepsis-Associated Immune Suppression and F-box Proteins, G966368916, G977340541
OpenAlex ID: W2991215660