Deep Sequencing of RNA from Blood and Oral Swab Samples Reveals the Presence of Nucleic Acid from a Number of Pathogens in Patients with Acute Ebola Virus Disease and Is Consistent with Bacterial Translocation across the Gut
Miles W. Carroll,
Sam Haldenby,
Natasha Y. Rickett,
Bernadett Pályi,
Isabel Garcia-Dorival,
Xuan Liu,
Gary Barker,
Joseph Akoi Bore,
Fara Raymond Koundouno,
E. Diane Williamson,
Thomas R. Laws,
Romy Kerber,
Daouda Sissoko,
Nóra Magyar,
Antonino Di Caro,
Mirella Biava,
Tom E. Fletcher,
Armand Sprecher,
Lisa F. P. Ng,
Laurent Rénia,
N’faly Magassouba,
Stephan Günther,
Roman Wölfel,
Kilian Stoecker,
David A. Matthews,
Julian A. Hiscox
Affiliations
Miles W. Carroll
Public Health England, Porton Down, Salisbury, United Kingdom
Sam Haldenby
Centre for Genomic Research Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom
Natasha Y. Rickett
Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom
Bernadett Pályi
National Public Health Institute, National Biosafety Laboratory, Budapest, Hungary
Isabel Garcia-Dorival
Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom
Xuan Liu
Centre for Genomic Research Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom
Gary Barker
School of Biological Sciences, University of Bristol, Bristol, United Kingdom
Joseph Akoi Bore
European Mobile Laboratory, Hamburg, Germany
Fara Raymond Koundouno
European Mobile Laboratory, Hamburg, Germany
E. Diane Williamson
Defence Science Technology Laboratories, Porton Down, United Kingdom
Thomas R. Laws
Defence Science Technology Laboratories, Porton Down, United Kingdom
Romy Kerber
Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany
Daouda Sissoko
Bordeaux Hospital University Center, INSERM U1219, Bordeaux University, Bordeaux, France
Nóra Magyar
National Public Health Institute, National Biosafety Laboratory, Budapest, Hungary
Antonino Di Caro
European Mobile Laboratory, Hamburg, Germany
Mirella Biava
European Mobile Laboratory, Hamburg, Germany
Tom E. Fletcher
Liverpool School of Tropical Medicine, Liverpool, United Kingdom
Armand Sprecher
Médecins Sans Frontières, Brussels, Belgium
Lisa F. P. Ng
Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom
Laurent Rénia
Singapore Immunology Network, Agency for Science, Technology and Research, Singapore
N’faly Magassouba
Université Gamal Abdel Nasser de Conakry, Laboratoire Des Fievres Hemorragiques en Guinee, Conakry, Guinea
Stephan Günther
European Mobile Laboratory, Hamburg, Germany
Roman Wölfel
Bundeswehr Institute of Microbiology, Munich, Germany
Kilian Stoecker
Bundeswehr Institute of Microbiology, Munich, Germany
David A. Matthews
School of Cellular and Molecular Medicine, University of Bristol, Bristol, United Kingdom
Julian A. Hiscox
Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom
ABSTRACT In this study, samples from the 2013–2016 West African Ebola virus outbreak from patients in Guinea with Ebola virus disease (EVD) were analyzed to discover and classify what other pathogens were present. Throat swabs were taken from deceased EVD patients, and peripheral blood samples were analyzed that had been taken from patients when they presented at the treatment center with acute illness. High-throughput RNA sequencing (RNA-seq) and bioinformatics were used to identify the potential microorganisms. This approach confirmed Ebola virus (EBOV) in all samples from patients diagnosed as acute positive for the virus by quantitative reverse transcription-PCR in deployed field laboratories. Nucleic acid mapping to Plasmodium was also used on the patient samples, confirming results obtained with an antigen-based rapid diagnostic test (RDT) conducted in the field laboratories. The data suggested that a high Plasmodium load, as determined by sequence read depth, was associated with mortality and influenced the host response, whereas a lower parasite load did not appear to affect outcome. The identifications of selected bacteria from throat swabs via RNA-seq were confirmed by culture. The data indicated that the potential pathogens identified in the blood samples were associated with translocation from the gut, suggesting the presence of bacteremia, which transcriptome data suggested may induce or aggravate the acute-phase response observed during EVD. Transcripts mapping to different viruses were also identified, including those indicative of lytic infections. The development of high-resolution analysis of samples from patients with EVD will help inform care pathways and the most appropriate general antimicrobial therapy to be used in a resource-poor setting. IMPORTANCE Our results highlight the identification of an array of pathogens in the blood of patients with Ebola virus disease (EVD). This has not been done before, and the data have important implications for the treatment of patients with EVD, particularly considering antibiotic stewardship. We show that EVD patients who were also infected with Plasmodium, particularly at higher loads, had more adverse outcomes than patients with lower levels of Plasmodium. However, the presence of Plasmodium did not influence the innate immune response, and it is likely that the presence of EBOV dominated this response. Several viruses other than EBOV were identified, and bacteria associated with sepsis were also identified. These findings were indicative of bacterial translocation across the gut during the acute phase of EVD.