{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,12]],"date-time":"2026-04-12T07:43:09Z","timestamp":1775979789834,"version":"3.50.1"},"reference-count":26,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2019,2,16]],"date-time":"2019-02-16T00:00:00Z","timestamp":1550275200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Following the largest Ebola virus disease outbreak from 2013 to 2016, viral RNA has been detected in survivors from semen and breast milk long after disease recovery. However, as there have been few cases of sexual transmission, it is unclear whether every RNA positive fluid sample contains infectious virus. Virus isolation, typically using cell culture or animal models, can serve as a tool to determine the infectivity of patient samples. However, the sensitivity of these methods has not been assessed for the Ebola virus isolate, Makona. Described here is an efficiency comparison of Ebola virus Makona isolation using Vero E6, Huh-7, monocyte-derived macrophage cells, and suckling laboratory mice. Isolation sensitivity was similar in all methods tested. Laboratory mice and Huh-7 cells were less affected by toxicity from breast milk than Vero E6 and MDM cells. However, the advantages associated with isolation in Huh-7 cells over laboratory mice, including cost effectiveness, sample volume preservation, and a reduction in animal use, make Huh-7 cells the preferred substrate tested for Ebola virus Makona isolation.<\/jats:p>","DOI":"10.3390\/v11020161","type":"journal-article","created":{"date-parts":[[2019,2,17]],"date-time":"2019-02-17T22:11:50Z","timestamp":1550441510000},"page":"161","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Ebola Virus Isolation Using Huh-7 Cells has Methodological Advantages and Similar Sensitivity to Isolation Using Other Cell Types and Suckling BALB\/c Laboratory Mice"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7410-9741","authenticated-orcid":false,"given":"James","family":"Logue","sequence":"first","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Walter","family":"Vargas Licona","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Timothy K.","family":"Cooper","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Becky","family":"Reeder","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Russel","family":"Byrum","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Qin","sequence":"additional","affiliation":[{"name":"Biostatistics Research Branch, National Institute of Allergy and Infectious Diseases, 5601 Fishers Lane, Rockville, MD 20852, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicole","family":"Deiuliis Murphy","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Cong","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8798-7714","authenticated-orcid":false,"given":"Amanda","family":"Bonilla","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jennifer","family":"Sword","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wade","family":"Weaver","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gregory","family":"Kocher","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gene G.","family":"Olinger","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter B.","family":"Jahrling","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"},{"name":"Emerging Viral Pathogens Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 21702, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lisa E.","family":"Hensley","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard S.","family":"Bennett","sequence":"additional","affiliation":[{"name":"Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 8200 Research Plaza, Frederick, MD 27102, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,2,16]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2018, December 20). Situation report: Ebola virus disease\u201410 June 2016. Available online: http:\/\/apps.who.int\/iris\/bitstream\/10665\/208883\/1\/ebolasitrep_10Jun2016_eng.pdf?ua=1."},{"key":"ref_2","unstructured":"Kuhn, J.H. (2017, November 08). Ebolavirus and marburgvirus infections. Available online: http:\/\/accessmedicine.mhmedical.com\/content.aspx?bookid=1130&sectionid=79739742."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1360","DOI":"10.1093\/cid\/ciw158","article-title":"Ebola virus disease complications as experienced by survivors in sierra leone","volume":"62","author":"Tiffany","year":"2016","journal-title":"Clin. Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1467","DOI":"10.1093\/infdis\/jiw079","article-title":"Ebola virus rna in the semen of male survivors of ebola virus disease: The uncertain gravitas of a privileged persistence","volume":"214","author":"Crozier","year":"2016","journal-title":"J. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Fischer, W., Brown, J., Wohl, D., Loftis, A., Tozay, S., Reeves, E., Pewu, K., Gorvego, G., Quellie, S., and Cunningham, C. (2017). Ebola vrus rna detection in semen more than two years after resolution of acute ebola virus infection. Open Forum Infect. Dis.","DOI":"10.1093\/ofid\/ofx155"},{"key":"ref_6","first-page":"479","article-title":"Possible sexual transmission of ebola virus\u2014Liberia, 2015","volume":"64","author":"Christie","year":"2015","journal-title":"MMWR Morb. Mortal. Wkly. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.1093\/cid\/ciw601","article-title":"Resurgence of ebola virus disease in guinea linked to a survivor with virus persistence in seminal fluid for more than 500 days","volume":"63","author":"Diallo","year":"2016","journal-title":"Clin. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2448","DOI":"10.1056\/NEJMoa1509773","article-title":"Molecular evidence of sexual transmission of ebola virus","volume":"373","author":"Mate","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_9","unstructured":"(2014). Liberia ebola Virus Disease Clinical Management Manual."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e736","DOI":"10.1016\/S2214-109X(16)30175-9","article-title":"Prevention of sexual transmission of ebola in liberia through a national semen testing and counselling programme for survivors: An analysis of ebola virus rna results and behavioural data","volume":"4","author":"Soka","year":"2016","journal-title":"Lancet Glob. Health"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e80","DOI":"10.1016\/S2214-109X(16)30243-1","article-title":"Persistence and clearance of ebola virus rna from seminal fluid of ebola virus disease survivors: A longitudinal analysis and modelling study","volume":"5","author":"Sissoko","year":"2017","journal-title":"The Lancet Global Health"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1128\/jcm.13.4.791-793.1981","article-title":"Plaque assay for ebola virus","volume":"13","author":"Moe","year":"1981","journal-title":"J. Clin. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1002\/jmv.1890040309","article-title":"Ebola virus virulence for newborn mice","volume":"4","author":"Jacob","year":"1979","journal-title":"J. Med. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1186\/1742-4690-7-55","article-title":"Semen-mediated enhancement of hiv infection is donor-dependent and correlates with the levels of sevi","volume":"7","author":"Kim","year":"2010","journal-title":"Retrovirology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1016\/j.cell.2007.10.014","article-title":"Semen-derived amyloid fibrils drastically enhance hiv infection","volume":"131","author":"Munch","year":"2007","journal-title":"Cell"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Cong, Y., Dyall, J., Hart, B.J., DeWald, L.E., Johnson, J.C., Postnikova, E., Zhou, H., Gross, R., Rojas, O., and Alexander, I. (2016). Evaluation of the activity of lamivudine and zidovudine against Ebola virus. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0166318"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e01331-14","DOI":"10.1128\/genomeA.01331-14","article-title":"Complete genome sequences of three ebola virus isolates from the 2014 outbreak in West Africa","volume":"2","author":"Hoenen","year":"2014","journal-title":"Genome Announc."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Schuit, M., Miller, D.M., Reddick-Elick, M.S., Wlazlowski, C.B., Filone, C.M., Herzog, A., Colf, L.A., Wahl-Jensen, V., Hevey, M., and Noah, J.W. (2016). Differences in the comparative stability of ebola virus makona-c05 and yambuku-mayinga in blood. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0148476"},{"key":"ref_19","first-page":"493","article-title":"A simple method of estimating fifty per cent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Hyg."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"5886","DOI":"10.1038\/s41598-017-06179-y","article-title":"High dose sertraline monotherapy fails to protect rhesus macaques from lethal challenge with ebola virus makona","volume":"7","author":"Honko","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1086\/515386","article-title":"A mouse model for evaluation of prophylaxis and therapy of ebola hemorrhagic fever","volume":"178","author":"Bray","year":"1998","journal-title":"J. Infect. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"7410","DOI":"10.1073\/pnas.1721646115","article-title":"Enhancement of ebola virus infection by seminal amyloid fibrils","volume":"115","author":"Bart","year":"2018","journal-title":"Proceedings of the National Academy of Sciences"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"S268","DOI":"10.1093\/infdis\/jiw145","article-title":"Ebola virus makona shows reduced lethality in an immune-deficient mouse model","volume":"214","author":"Smither","year":"2016","journal-title":"J. Infect. Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"S129","DOI":"10.1093\/infdis\/jiu681","article-title":"An improved reverse genetics system to overcome cell-type-dependent ebola virus genome plasticity","volume":"212","author":"Tsuda","year":"2015","journal-title":"J. Infect. Dis."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"JVI-00392","DOI":"10.1128\/JVI.00392-17","article-title":"Spontaneous mutation at amino acid 544 of the ebola virus glycoprotein potentiates virus entry and selection in tissue culture","volume":"91","author":"Ruedas","year":"2017","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"JVI-00177","DOI":"10.1128\/JVI.00177-17","article-title":"A polymorphism within the internal fusion loop of the ebola virus glycoprotein modulates host cell entry","volume":"91","author":"Hoffmann","year":"2017","journal-title":"J. Virol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/2\/161\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:32:40Z","timestamp":1760185960000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/2\/161"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,2,16]]},"references-count":26,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2019,2]]}},"alternative-id":["v11020161"],"URL":"https:\/\/doi.org\/10.3390\/v11020161","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,2,16]]}}}