title: Persistence and clearance of Ebola virus RNA from seminal fluid of Ebola virus disease survivors: a longitudinal analysis and modelling study.
abstract: By January, 2016, all known transmission chains of the Ebola virus disease (EVD) outbreak in west Africa had been stopped. However, there is concern about persistence of Ebola virus in the reproductive tract of men who have survived EVD. We aimed to use biostatistical modelling to describe the dynamics of Ebola virus RNA load in seminal fluid, including clearance parameters.
  
  
  
  In this longitudinal study, we recruited men who had been discharged from three Ebola treatment units in Guinea between January and July, 2015. Participants provided samples of seminal fluid at follow-up every 3-6 weeks, which we tested for Ebola virus RNA using quantitative real-time RT-PCR. Representative specimens from eight participants were then inoculated into immunodeficient mice to test for infectivity. We used a linear mixed-effect model to analyse the dynamics of virus persistence in seminal fluid over time.
  
  
  
  We enrolled 26 participants and tested 130 seminal fluid specimens; median follow up was 197 days (IQR 187-209 days) after enrolment, which corresponded to 255 days (228-287) after disease onset. Ebola virus RNA was detected in 86 semen specimens from 19 (73%) participants. Median duration of Ebola virus RNA detection was 158 days after onset (73-181; maximum 407 days at end of follow-up). Mathematical modelling of the quantitative time-series data showed a mean clearance rate of Ebola virus RNA from seminal fluid of -0·58 log units per month, although the clearance kinetic varied greatly between participants. Using our biostatistical model, we predict that 50% and 90% of male survivors clear Ebola virus RNA from seminal fluid at 115 days (90% prediction interval 72-160) and 294 days (212-399) after disease onset, respectively. We also predicted that the number of men positive for Ebola virus RNA in affected countries would decrease from about 50 in January 2016, to fewer than 1 person by July, 2016. Infectious virus was detected in 15 of 26 (58%) specimens tested in mice.
  
  
  
  Time to clearance of Ebola virus RNA from seminal fluid varies greatly between individuals and could be more than 13 months. Our predictions will assist in decision-making about surveillance and preventive measures in EVD outbreaks.
  
  
  
  This study was funded by European Union's Horizon 2020 research and innovation programme, Directorate-General for International Cooperation and Development of the European Commission, Institut national de la santé et de la recherche médicale (INSERM), German Research Foundation (DFG), and Innovative Medicines Initiative 2 Joint Undertaking.
authors.affiliation: INSERM U1219, Bordeaux University, Bordeaux, France; Bordeaux University Hospital, Bordeaux, France.
authors.forename: Daouda
authors.initials: D
authors.lastname: Sissoko
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany.
authors.forename: Sophie
authors.initials: S
authors.lastname: Duraffour
authors.affiliation: Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany; German Centre for Infection Research (DZIF), Partner Site Hamburg, Conakry, Guinea.
authors.forename: Romy
authors.initials: R
authors.lastname: Kerber
authors.affiliation: Centre de Formation et de Recherche en Santé Rurale de Mafèrinyah, Conakry, Guinea.
authors.forename: Jacques Seraphin
authors.initials: JS
authors.lastname: Kolie
authors.affiliation: Centre de Formation et de Recherche en Santé Rurale de Mafèrinyah, Conakry, Guinea.
authors.forename: Abdoul Habib
authors.initials: AH
authors.lastname: Beavogui
authors.affiliation: Department of Infectious Diseases, Donka Hospital, Conakry, Guinea.
authors.forename: Alseny-Modet
authors.initials: AM
authors.lastname: Camara
authors.affiliation: INSERM U1219, Bordeaux University, Bordeaux, France; PAC-CI, ANRS Research Site, Treichville University Hospital, Abidjan, Côte d'Ivoire.
authors.forename: Géraldine
authors.initials: G
authors.lastname: Colin
authors.affiliation: Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany; German Centre for Infection Research (DZIF), Partner Site Hamburg, Conakry, Guinea.
authors.forename: Toni
authors.initials: T
authors.lastname: Rieger
authors.affiliation: Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany; German Centre for Infection Research (DZIF), Partner Site Hamburg, Conakry, Guinea.
authors.forename: Lisa
authors.initials: L
authors.lastname: Oestereich
authors.affiliation: National Centre for Epidemiology, Hungarian National Biosafety Laboratory, Budapest, Hungary.
authors.forename: Bernadett
authors.initials: B
authors.lastname: Pályi
authors.affiliation: Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany; German Centre for Infection Research (DZIF), Partner Site Hamburg, Conakry, Guinea.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Wurr
authors.affiliation: INSERM, IAME, UMR 1137 Paris, and Université Paris Diderot, Sorbonne Paris Cité, France.
authors.forename: Jeremie
authors.initials: J
authors.lastname: Guedj
authors.affiliation: INSERM, IAME, UMR 1137 Paris, and Université Paris Diderot, Sorbonne Paris Cité, France.
authors.forename: Thi Huyen Tram
authors.initials: THT
authors.lastname: Nguyen
authors.affiliation: Department of Infectious Disease Epidemiology, London School of Hygiene & Tropical Medicine, London, UK.
authors.forename: Rosalind M
authors.initials: RM
authors.lastname: Eggo
authors.affiliation: Department of Infectious Disease Epidemiology, London School of Hygiene & Tropical Medicine, London, UK.
authors.forename: Conall H
authors.initials: CH
authors.lastname: Watson
authors.affiliation: Department of Infectious Disease Epidemiology, London School of Hygiene & Tropical Medicine, London, UK.
authors.forename: W John
authors.initials: WJ
authors.lastname: Edmunds
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Ministry of Health Guinea, Conakry, Guinea.
authors.forename: Joseph Akoi
authors.initials: JA
authors.lastname: Bore
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Ministry of Health Guinea, Conakry, Guinea.
authors.forename: Fara Raymond
authors.initials: FR
authors.lastname: Koundouno
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany.
authors.forename: Mar
authors.initials: M
authors.lastname: Cabeza-Cabrerizo
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; University College London, London, UK.
authors.forename: Lisa L
authors.initials: LL
authors.lastname: Carter
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Laboratory for Clinical and Epidemiological Virology, Department of Microbiology and Immunology, KU Leuven, Leuven, Belgium.
authors.forename: Liana Eleni
authors.initials: LE
authors.lastname: Kafetzopoulou
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Public Health England, Porton Down, Salisbury, UK.
authors.forename: Eeva
authors.initials: E
authors.lastname: Kuisma
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Robert Koch Institute, Berlin, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Michel
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany.
authors.forename: Livia Victoria
authors.initials: LV
authors.lastname: Patrono
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Institute of Infection and Global Health, University of Liverpool, Liverpool, UK.
authors.forename: Natasha Y
authors.initials: NY
authors.lastname: Rickett
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Institute of Virology, Technische Universität München, Munich, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Singethan
authors.affiliation: Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany; German Centre for Infection Research (DZIF), Partner Site Hamburg, Conakry, Guinea.
authors.forename: Martin
authors.initials: M
authors.lastname: Rudolf
authors.affiliation: Robert Koch Institute, Berlin, Germany.
authors.forename: Angelika
authors.initials: A
authors.lastname: Lander
authors.affiliation: Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany; German Centre for Infection Research (DZIF), Partner Site Hamburg, Conakry, Guinea.
authors.forename: Elisa
authors.initials: E
authors.lastname: Pallasch
authors.affiliation: Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany; German Centre for Infection Research (DZIF), Partner Site Hamburg, Conakry, Guinea.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Bockholt
authors.affiliation: German Centre for Infection Research (DZIF), Partner Site Hamburg, Conakry, Guinea; Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg.
authors.forename: Estefanía
authors.initials: E
authors.lastname: Rodríguez
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; National Institute for Infectious Diseases Lazzaro Spallanzani, Rome, Italy.
authors.forename: Antonino
authors.initials: A
authors.lastname: Di Caro
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Bundeswehr Institute of Microbiology, Munich, Switzerland.
authors.forename: Roman
authors.initials: R
authors.lastname: Wölfel
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Gabriel
authors.affiliation: WHO, Geneva, Switzerland.
authors.forename: Céline
authors.initials: C
authors.lastname: Gurry
authors.affiliation: WHO, Geneva, Switzerland.
authors.forename: Pierre
authors.initials: P
authors.lastname: Formenty
authors.affiliation: Ministry of Health Guinea, Conakry, Guinea.
authors.forename: Sakoba
authors.initials: S
authors.lastname: Keïta
authors.affiliation: INSERM U1219, Bordeaux University, Bordeaux, France; Bordeaux University Hospital, Bordeaux, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Malvy
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Public Health England, Porton Down, Salisbury, UK; University of Southampton, South General Hospital, Southampton, UK.
authors.forename: Miles W
authors.initials: MW
authors.lastname: Carroll
authors.affiliation: INSERM U1219, Bordeaux University, Bordeaux, France; PAC-CI, ANRS Research Site, Treichville University Hospital, Abidjan, Côte d'Ivoire.
authors.forename: Xavier
authors.initials: X
authors.lastname: Anglaret
authors.affiliation: The European Mobile Laboratory Consortium, Hamburg, Germany; Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany; German Centre for Infection Research (DZIF), Partner Site Hamburg, Conakry, Guinea. Electronic address: guenther@bni.uni-hamburg.de.
authors.forename: Stephan
authors.initials: S
authors.lastname: Günther
mesh_terms: D000328:Adult; D029043:Ebolavirus / Q000235:genetics / Q000302:isolation & purification*; D016701:Guinea; D019142:Hemorrhagic Fever, Ebola / Q000453:epidemiology / Q000517:prevention & control* / Q000635:transmission* / Q000821:virology; D006801:Humans; D008137:Longitudinal Studies; D008297:Male; D008875:Middle Aged; D015233:Models, Statistical; D012313:RNA; D012661:Semen; D017741:Survivors; D013997:Time Factors
chemical_list: D012313:RNA