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1.
Microbial volatiles as diagnostic biomarkers of bacterial lung infection in mechanically ventilated patients
Waqar M Ahmed, Dominic Fenn, Iain R. White, Breanna Dixon, Tamara M E Nijsen, Hugo H Knobel, Paul Brinkman, Pouline M P van Oort, Marcus J Schultz, Paul Dark, Royston Goodacre, Timothy Felton, Lieuwe D J Bos, Stephen J. Fowler, 2022, original scientific article

Abstract: Background Early and accurate recognition of respiratory pathogens is crucial to prevent increased risk of mortality in critically ill patients. Microbial-derived volatile organic compounds (mVOCs) in exhaled breath could be used as non-invasive biomarkers of infection to support clinical diagnosis. Methods In this study, we investigated the diagnostic potential of in vitro confirmed mVOCs in the exhaled breath of patients under mechanically ventilation from the BreathDx study. Samples were analysed by thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). Results Pathogens from bronchoalveolar lavage (BAL) cultures were identified in 45/89 patients and S. aureus was the most commonly identified pathogen (n = 15). Out of 19 mVOCs detected in the in vitro culture headspace of four common respiratory pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli), 14 were found in exhaled breath samples. Higher concentrations of two mVOCs were found in the exhaled breath of patients infected with S. aureus compared to those without (3-methylbutanal p < 0.01. AUROC = 0.81-0.87 and 3-methylbutanoic acid p = 0.01. AUROC = 0.79-0.80). In addition, bacteria identified from BAL cultures which are known to metabolise tryptophan (Escherichia coli, Klebsiella oxytoca and Haemophilus influenzae) were grouped and found to produce higher concentrations of indole compared to breath samples with culture-negative (p = 0.034) and other pathogen-positive (p = 0.049) samples. Conclusions This study demonstrates the capability of using mVOCs to detect the presence of specific pathogen groups with potential to support clinical diagnosis. Although not all mVOCs were found in patient samples within this small pilot study, further targeted and qualitative investigation is warranted using multi-centre clinical studies.
Keywords: Breath, VOCs, infection, respiratory pathogens, VAP
Published in RUNG: 28.11.2022; Views: 2172; Downloads: 0
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2.
Composition and diversity analysis of the lung microbiome in patients with suspected ventilator-associated pneumonia
Dominic Fenn, Mahmoud Abdel-Aziz, Pouline van Oort, Paul Brinkman, Waqar Ahmed, Timothy Felton, Antonio Artigas, Pedro Póvoa, Ignacio Martin-Loeches, Marcus Schultz, Paul Dark, Stephen Fowler, Iain R. White, 2022, original scientific article

Abstract: Background: Ventilator-associated pneumonia (VAP) is associated with high morbidity and health care costs, yet diagnosis remains a challenge. Analysis of airway microbiota by amplicon sequencing provides a possible solution, as pneumonia is characterised by a disruption of the microbiome. However, studies evaluating the diagnostic capabilities of microbiome analysis are limited, with a lack of alignment on possible biomarkers. Using bronchoalveolar lavage fluid (BALF) from ventilated adult patients suspected of VAP, we aimed to explore how key characteristics of the microbiome differ between patients with positive and negative BALF cultures and whether any differences could have a clinically relevant role. Methods: BALF from patients suspected of VAP was analysed using 16s rRNA sequencing in order to: (1) differentiate between patients with and without a positive culture; (2) determine if there was any association between microbiome diversity and local inflammatory response; and (3) correctly identify pathogens detected by conventional culture. Results: Thirty-seven of 90 ICU patients with suspected VAP had positive cultures. Patients with a positive culture had significant microbiome dysbiosis with reduced alpha diversity. However, gross compositional variance was not strongly associated with culture positivity (AUROCC range 0.66–0.71). Patients with a positive culture had a significantly higher relative abundance of pathogenic bacteria compared to those without [0.45 (IQR 0.10–0.84), 0.02 (IQR 0.004–0.09), respectively], and an increased interleukin (IL)-1β was associated with reduced species evenness (rs = − 0.33, p < 0.01) and increased pathogenic bacteria presence (rs = 0.28, p = 0.013). Untargeted 16s rRNA pathogen detection was limited by false positives, while the use of pathogen-specific relative abundance thresholds showed better diagnostic accuracy (AUROCC range 0.89–0.998). Conclusion: Patients with positive BALF culture had increased dysbiosis and genus dominance. An increased caspase-1-dependent IL-1b expression was associated with a reduced species evenness and increased pathogenic bacterial presence, providing a possible causal link between microbiome dysbiosis and lung injury development in VAP. However, measures of diversity were an unreliable predictor of culture positivity and 16s sequencing used agnostically could not usefully identify pathogens; this could be overcome if pathogen-specific relative abundance thresholds are used.
Keywords: Microbiome, Next-generation sequencing, Ventilator-associated pneumonia
Published in RUNG: 26.10.2022; Views: 1905; Downloads: 0
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