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      A 40-Month Follow-Up of Ebola Virus Disease Survivors in Guinea (PostEbogui) Reveals Long-Term Detection of Ebola Viral Ribonucleic Acid in Semen and Breast Milk

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          Abstract

          Background

          With the increasing frequency and impact of Ebola virus disease (EVD) outbreaks illustrated by recent epidemics, a good understanding of the extent of viral persistance or ribonucleic acid (RNA) detection in body fluids from survivors is urgently needed.

          Methods

          Ebola viral RNA shedding was studied with molecular assays in semen (n = 1368), urine (n = 1875), cervicovaginal fluid (n = 549), saliva (n = 900), breast milk (n = 168), and feces (n = 558) from EVD survivors in Guinea (PostEbogui cohort, n = 802) at a regular base period until 40 months after inclusion.

          Results

          Twenty-seven of 277 (9.8%) male survivors tested positive for Ebola RNA in at least 1 semen sample. The probability of remaining positive for Ebola RNA in semen was estimated at 93.02% and 60.12% after 3 and 6 months. Viral RNA in semen was more frequent in patients with eye pain ( P = .036), joint pain ( P = .047), and higher antibody levels to Ebola virus antigens (nucleoprotein [ P = .001], glycoprotein [ P = .05], and viral protein-40 [ P = .05]). Ebola RNA was only rarely detected in the following body fluids from EVD survivors: saliva (1 of 454), urine (2 of 593), breast milk (2 of 168), cervicovaginal secretions (0 of 273), and feces (0 of 330). Ribonucleic acid was detected in breast milk 1 month after delivery but 500 days after discharge of Ebola treatment unit (ETU) in 1 woman who became pregnant 7 months after discharge from the ETU.

          Conclusions

          The frequency and potential long-term presence of viral RNA in semen confirmed that systematic prevention measures in male survivors are required. Our observation in breast milk suggests that our knowledge on viral reservoir in immune-privileged sites and its impact are still incomplete.

          Abstract

          On 5,400 body fluid samples from Ebola Virus Disease (EVD) survivors, during 40 months follow-up in Guinea, RNA was observed in semen and breast milk for up to 500 days, illustrating the complexity of the viral reservoir and management of survivors.

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          Most cited references30

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          Ebola RNA Persistence in Semen of Ebola Virus Disease Survivors - Preliminary Report.

          Background Ebola virus has been detected in the semen of men after their recovery from Ebola virus disease (EVD), but little information is available about its prevalence or the duration of its persistence. We report the initial findings of a pilot study involving survivors of EVD in Sierra Leone. Methods We enrolled a convenience sample of 100 male survivors of EVD in Sierra Leone, at different times after their recovery from EVD, and recorded self-reported information about sociodemographic characteristics, the EVD episode, and health status. Semen specimens obtained at baseline were tested by means of a quantitative reverse-transcriptase-polymerase-chain-reaction (RT-PCR) assay with the use of the target-gene sequences of NP and VP40. Results A total of 93 participants provided an initial semen specimen for analysis, of whom 46 (49%) had positive results on quantitative RT-PCR. Ebola virus RNA was detected in the semen of all 9 men who had a specimen obtained 2 to 3 months after the onset of EVD, in the semen of 26 of 40 (65%) who had a specimen obtained 4 to 6 months after onset, and in the semen of 11 of 43 (26%) who had a specimen obtained 7 to 9 months after onset; the results for 1 participant who had a specimen obtained at 10 months were indeterminate. The median cycle-threshold values (for which higher values indicate lower RNA levels) were 32.0 with the NP gene target and 31.1 with the VP40 gene target for specimens obtained at 2 to 3 months, 34.5 and 32.3, respectively, for specimens obtained at 4 to 6 months, and 37.0 and 35.6, respectively, for specimens obtained at 7 to 9 months. Conclusions These data showed the persistence of Ebola virus RNA in semen and declining persistence with increasing months since the onset of EVD. We do not yet have data on the extent to which positivity on RT-PCR is associated with virus infectivity. Although cases of suspected sexual transmission of Ebola have been reported, they are rare; hence the risk of sexual transmission of the Ebola virus is being investigated. (Funded by the World Health Organization and others.).
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            Persistence and genetic stability of Ebola virus during the outbreak in Kikwit, Democratic Republic of the Congo, 1995.

            Ebola virus persistence was examined in body fluids from 12 convalescent patients by virus isolation and reverse transcription-polymerase chain reaction (RT-PCR) during the 1995 Ebola hemorrhagic fever outbreak in Kikwit, Democratic Republic of the Congo. Virus RNA could be detected for up to 33 days in vaginal, rectal, and conjunctival swabs of 1 patient and up to 101 days in the seminal fluid of 4 patients. Infectious virus was detected in 1 seminal fluid sample obtained 82 days after disease onset. Sequence analysis of an RT-PCR fragment of the most variable region of the glycoprotein gene amplified from 9 patients revealed no nucleotide changes. The patient samples were selected so that they would include some from a suspected line of transmission with at least three human-to-human passages, some from 5 survivors and 4 deceased patients, and 2 from patients who provided multiple samples through convalescence. There was no evidence of different virus variants cocirculating during the outbreak or of genetic variation accumulating during human-to-human passage or during prolonged persistence in individual patients.
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              Resurgence of Ebola Virus Disease in Guinea Linked to a Survivor With Virus Persistence in Seminal Fluid for More Than 500 Days

              We report on an Ebola virus disease (EVD) survivor who showed Ebola virus in seminal fluid 531 days after onset of disease. The persisting virus was sexually transmitted in February 2016, about 470 days after onset of symptoms, and caused a new cluster of EVD in Guinea and Liberia.
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                Author and article information

                Journal
                Open Forum Infect Dis
                Open Forum Infect Dis
                ofid
                Open Forum Infectious Diseases
                Oxford University Press (US )
                2328-8957
                December 2019
                08 November 2019
                08 November 2019
                : 6
                : 12
                : ofz482
                Affiliations
                [1 ] TransVIHMI, IRD/INSERM/Monpellier University , Montpellier, France
                [2 ] Centre de Recherche et de Formation en Infectiologie de Guinée, Université Gamal Abdel Nasser de Conakry , Conakry, Guinea
                [3 ] Institut National de Santé Publique , Conakry, Guinea
                [4 ] Laboratory of Virology, Conakry University, Projet de Recherche sur les Fièvres Hémorragiques en Guinée , Conakry, Guinea
                [5 ] Institut Pasteur, Unit of Biology of Emerging Viral Infections - National Reference Center for Viral Hemorrhagic Fevers , Lyon, France
                [6 ] Centre International de Recherche en Infectiologie, Lyon University - INSERM - ENS Lyon - CNRS , Lyon, France
                [7 ] Laboratoire P4 Inserm-Jean Mérieux , Lyon, France
                [8 ] Hospices Civils de Lyon, Service de Biostatistique , Lyon, France
                [9 ] CNRS UMR 5558 Laboratoire de Biométrie et Biologie Évolutive, Équipe Biostatistique-Santé , Villeurbanne, France
                [10 ] Donka National Hospital , Conakry, Guinea
                [11 ] IRD/INSERM/Monpellier University , Montpellier, France
                [12 ] Pasteur Institute/Unit of Biology of Emerging Viral Infections , Lyon, France
                [15 ] Forecariah Prefectoral Hospital , Forecariah, Guinea
                [16 ] Reacting, INSERM , Paris, France
                [17 ] Macenta Prefectoral Hospital , Macenta, Guinea
                [18 ] Ministry of Health , Conakry, Guinea
                [19 ] Institut National de la Santé Publique , Conakry, Guinea
                [20 ] INSERM , Paris, France
                [21 ] Laboratory Virology of University Conakry Projet de Recherche sur les Fièvres Hémorragiques en Guinée , Conakry, Guinea
                [22 ] ALIMA, Conakry , Guinea
                [23 ] Monpellier University , Montpellier, France
                [24 ] Laboratoire P4 Inserm-Jean Mérieux , Inserm, Lyon, France
                [25 ] N’Zérékoré Regional Hospital , N’Zérékoré, Guinea
                Author notes

                A. K. K. and N. V. contributed equally to this work.

                Correspondence: Martine Peeters, PhD, IRD, TransVIHMI, 911 avenue Agropolis, 34394 Montpellier Cedex5, France ( martine.peeters@ 123456ird.fr ).
                Author information
                http://orcid.org/0000-0001-7022-2643
                http://orcid.org/0000-0001-6738-8730
                Article
                ofz482
                10.1093/ofid/ofz482
                7047953
                32128327
                96a872d6-b787-43d9-a3c1-10fe6b2a0f74
                © The Author(s) 2019. Published by Oxford University Press on behalf of Infectious Diseases Society of America.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence ( http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

                History
                : 26 September 2019
                : 26 October 2019
                : 05 November 2019
                Page count
                Pages: 8
                Funding
                Funded by: Institut National de la Santé et de la Recherche Médicale/Ebola Task Force
                Funded by: REACTing
                Funded by: Institut de Recherche pour le Développement 10.13039/100012947
                Funded by: Montpellier Université d’Excellence
                Award ID: ANR-16-IDEX-0006
                Categories
                Major Article
                Editor's Choice

                body fluids,breast milk,ebola,guinea,semen
                body fluids, breast milk, ebola, guinea, semen

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