Detection of Bacterial Pathogens from Broncho-Alveolar Lavage by Next-Generation Sequencing

Leo, Stefano; Gaïa, Nadia; Ruppé, Etienne; Emonet, Stephane; Girard, Myriam; Lazarevic, Vladimir; Schrenzel, Jacques

doi:10.3390/ijms18092011

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Detection of Bacterial Pathogens from Broncho-Alveolar Lavage by Next-Generation Sequencing

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Genomic Research Laboratory, Service of Infectious Diseases, Geneva University Hospitals (HUG), 1205 Geneva, Switzerland

Service of Infectious Diseases, Geneva University Hospitals and Faculty of Medicine, 1205 Geneva, Switzerland

Bacteriology Laboratory, Geneva University Hospitals and Faculty of Medicine, 1205 Geneva, Switzerland

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2017, 18(9), 2011; https://doi.org/10.3390/ijms18092011

Received: 15 August 2017 / Revised: 10 September 2017 / Accepted: 13 September 2017 / Published: 20 September 2017

(This article belongs to the Special Issue Deciphering the Human Microbiota: Methods and Impact on Human Health)

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Abstract

The applications of whole-metagenome shotgun sequencing (WMGS) in routine clinical analysis are still limited. A combination of a DNA extraction procedure, sequencing, and bioinformatics tools is essential for the removal of human DNA and for improving bacterial species identification in a timely manner. We tackled these issues with a broncho-alveolar lavage (BAL) sample from an immunocompromised patient who had developed severe chronic pneumonia. We extracted DNA from the BAL sample with protocols based either on sequential lysis of human and bacterial cells or on the mechanical disruption of all cells. Metagenomic libraries were sequenced on Illumina HiSeq platforms. Microbial community composition was determined by k-mer analysis or by mapping to taxonomic markers. Results were compared to those obtained by conventional clinical culture and molecular methods. Compared to mechanical cell disruption, a sequential lysis protocol resulted in a significantly increased proportion of bacterial DNA over human DNA and higher sequence coverage of Mycobacterium abscessus, Corynebacterium jeikeium and Rothia dentocariosa, the bacteria reported by clinical microbiology tests. In addition, we identified anaerobic bacteria not searched for by the clinical laboratory. Our results further support the implementation of WMGS in clinical routine diagnosis for bacterial identification. View Full-Text

Keywords: whole-metagenome shotgun; clinical metagenomics; broncho-alveolar lavage

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MDPI and ACS Style

Leo, S.; Gaïa, N.; Ruppé, E.; Emonet, S.; Girard, M.; Lazarevic, V.; Schrenzel, J. Detection of Bacterial Pathogens from Broncho-Alveolar Lavage by Next-Generation Sequencing. Int. J. Mol. Sci. 2017, 18, 2011. https://doi.org/10.3390/ijms18092011

AMA Style

Leo S, Gaïa N, Ruppé E, Emonet S, Girard M, Lazarevic V, Schrenzel J. Detection of Bacterial Pathogens from Broncho-Alveolar Lavage by Next-Generation Sequencing. International Journal of Molecular Sciences. 2017; 18(9):2011. https://doi.org/10.3390/ijms18092011

Chicago/Turabian Style

Leo, Stefano, Nadia Gaïa, Etienne Ruppé, Stephane Emonet, Myriam Girard, Vladimir Lazarevic, and Jacques Schrenzel. 2017. "Detection of Bacterial Pathogens from Broncho-Alveolar Lavage by Next-Generation Sequencing" International Journal of Molecular Sciences 18, no. 9: 2011. https://doi.org/10.3390/ijms18092011

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Detection of Bacterial Pathogens from Broncho-Alveolar Lavage by Next-Generation Sequencing

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