Ebola Virus Persistence in Semen of Male Survivors

Uyeki, Timothy M.; Erickson, Bobbie R.; Brown, Shelley; McElroy, Anita K.; Cannon, Deborah; Gibbons, Aridth; Sealy, Tara K.; Kainulainen, Markus H.; Schuh, Amy J.; Kraft, Colleen S.; Mehta, Aneesh K.; Lyon, G. Marshall; Varkey, Jay B.; Ribner, Bruce S.; Ellison, Richard T.; Carmody, Ellie; Nau, Gerard J.; Spiropoulou, Christina F.; Nichol, Stuart T.; Ströher, Ute

doi:10.1093/cid/ciw202

Cited by 98 publications

(79 citation statements)

References 8 publications

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“…The different levels of susceptibility to RNase digestion suggest possible differences between vRNP and cRNP with respect to their structures or compositions. Since vRNP exists in the EBOV virion structure, the extraordinary stability of vRNP may reflect the astounding persistence of EBOV virions or genomes in the tissues of convalescent patients (35)(36)(37) or in environment settings (38)(39)(40)(41). As EBOV persistence has increasingly become a research focus (42), our replicon model may provide a useful tool to analyze the contributions of the vRNP stability to persistent EBOV infection.…”

Section: Discussionmentioning

confidence: 99%

Novel Stable Ebola Virus Minigenome Replicon Reveals Remarkable Stability of the Viral Genome

Tao

Gan

Guo

et al. 2017

J Virol

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Ebola virus (EBOV) causes severe hemorrhagic fever in humans and other primates with a high case fatality rate. No approved drug or vaccine of EBOV is available, which necessitates better understanding of the virus life cycle. Studies on EBOV have been hampered because experimentations involving live virus are restricted to biosafety level 4 (BSL4) laboratories. The EBOV minigenome system has provided researchers with the opportunity to study EBOV under BSL2 conditions. Here, we developed a novel EBOV minigenome replicon which, to our knowledge, is the first EBOV cell culture system that can stably replicate and transcribe the EBOV minigenome. The minigenomic RNA harboring a Gaussia luciferase and hygromycinresistant marker can replicate for months in a helper cell stably expressing viral nucleoprotein (NP), viral protein 35 (VP35), VP30, and L proteins. Quantification of viral RNA (vRNA), cRNA, and mRNA levels of the EBOV minigenome demonstrated that the stable EBOV replicon had much-more-active minigenome replication than previously developed transient-transfection-based EBOV minigenome systems, which recapitulate viral primary transcription more than genome replication. Interestingly, minigenome replication in the stable EBOV replicon cells was insensitive to interferon treatment or RNA interference. Moreover, RNase digestion of the replicon cell lysates revealed the remarkably stable nature of the EBOV minigenomic vRNA ribonucleoprotein complex, which may help improve understanding of EBOV persistence in convalescent patients. IMPORTANCEThe scope and severity of the recent Ebola outbreak in Western Africa justified a more comprehensive investigation of the causative risk group 4 agent Ebola virus (EBOV). Study of EBOV replication and antiviral development can be facilitated by developing a cell culture system that allows experimentation under biosafety level 2 conditions. Here, we developed a novel stable EBOV minigenome replicon which, to our knowledge, is the first EBOV cell culture system that can stably replicate and transcribe the EBOV minigenome. The replicon system had more-active genome replication than previously developed transient-transfection-based EBOV minigenome systems, providing a convenient surrogate system to study EBOV replication. Furthermore, self-replicating minigenomic vRNA in the replicon cells displayed strong stability in response to interferon treatment, RNA silencing, and RNase digestion, which may provide an explanation for the persistence of EBOV in survivors.

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Section: Discussionmentioning

confidence: 99%

Novel Stable Ebola Virus Minigenome Replicon Reveals Remarkable Stability of the Viral Genome

Tao

Gan

Guo

et al. 2017

J Virol

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“…An expanded window for diagnosis in CHIKV is also needed since IgM seropositivity may persist for months after the initial acute infection. Many other viruses have been detected in semen, such as Ebola, HIV, HBV, HPV, HSV-1, HSV-2, Marburg, but there has been no published data so far on the presence of CHIKV in semen [13], [14], [15], [16], [17].…”

Section: Introductionmentioning

confidence: 99%

Prolonged shedding of Chikungunya virus in semen and urine: A new perspective for diagnosis and implications for transmission

Bandeira

Campos

Rocha³

et al. 2016

IDCases

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AbsractWe report the presence of Chikungunya (CHIKV) RNA in the blood, urine and semen during the acute phase of the disease in an adult with a dual infection with Dengue virus type 3. The patient, a 25 yr-old man from Salvador, Brazil, reported a 6-day duration of high fever, arthralgia, myalgia, headache and photophobia.Blood and semen specimens were positive for CHIKV in the first collected samples; semen and urine specimens were positive for CHIKV after 30 days of symptoms onset. DENV-3 RNA was positive in blood specimen when first collected 6 days after the initiation of symptoms.We describe for the first time the presence of CHIKV RNA in urine and semen for an extended period of time and we address the possible implications of these findings for diagnosis and transmission dynamics.

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“…Though many individuals recovered from disease, clear evidence demonstrates that Ebola virus (EBOV) can persist in EVD survivors long after signs and symptoms have subsided (MacDermott and Bausch, 2016; World Health Organization, 2016a). Persistent genomic virus RNA signals have been detected in EVD survivors over a year after resolution of both clinical disease and viremia(Crozier, 2016; Deen et al, 2017; Uyeki et al, 2016). As a result, sexual and vertical transmission of the virus may occur, as evidenced by at least 3 EVD clusters in Liberia, Sierra Leone, and Guinea (Arias et al, 2016; Blackley et al, 2016;Fischer et al, 2016; Fischer and Wohl, 2016; Mate et al, 2015; Sow et al, 2016; Yang et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Use of the Filovirus Animal Non-Clinical Group (FANG) Ebola virus immuno-assay requires fewer study participants to power a study than the Alpha Diagnostic International assay

Logue

Tuznik

Follmann

et al. 2018

Journal of Virological Methods

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As part of the scientific community’s development of medical countermeasures against Ebola virus disease, optimization of standardized assays for product evaluation is paramount. The recent outbreak heightened awareness to the scarcity of available assays and limited information on performance and reproducibility. To evaluate the immunogenicity of vaccines entering Phase I–III trials and to identify survivors, two enzyme-linked immunosorbent assays, the Filovirus Animal Non-Clinical Group assay and the Alpha Diagnostics International assay, were evaluated for detection of immunoglobulin G against Ebola virus glycoprotein. We found that the Filovirus Animal Nonclinical Group assay produced a wider range of relative antibody concentrations, higher assay precision, larger relative accuracy range, and lower regional background. Additionally, to sufficiently power a vaccine trial, use of the Filovirus Animal Nonclinical Group assay would require one third the number of participants than the Alpha Diagnostics International assay. This reduction in needed study participants will require less money, fewer man hours, and much less time to evaluate vaccine immunogenicity.

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Ebola Virus Persistence in Semen of Male Survivors

Abstract: We investigated the duration of Ebola virus (EBOV) RNA and infectious EBOV in semen specimens of 5 Ebola virus disease (EVD) survivors. EBOV RNA and infectious EBOV was detected by real-time RT-PCR and virus culture out to 290 days and 70 days, respectively, after EVD onset.

Cited by 98 publications

References 8 publications

Novel Stable Ebola Virus Minigenome Replicon Reveals Remarkable Stability of the Viral Genome

Novel Stable Ebola Virus Minigenome Replicon Reveals Remarkable Stability of the Viral Genome

Prolonged shedding of Chikungunya virus in semen and urine: A new perspective for diagnosis and implications for transmission

Use of the Filovirus Animal Non-Clinical Group (FANG) Ebola virus immuno-assay requires fewer study participants to power a study than the Alpha Diagnostic International assay

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