DOCSLIB.ORG

Ebola Virus Disease

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Ebola Virus Disease PRIMER Ebola virus disease Shevin T. Jacob1,2, Ian Crozier3, William A. Fischer II4, Angela Hewlett5, Colleen S. Kraft6, Marc-Antoine de La Vega7, Moses J. Soka8, Victoria Wahl9, Anthony Griffiths10, Laura Bollinger11 and Jens H. Kuhn11 ✉ Abstract | Ebola virus disease (EVD) is a severe and frequently lethal disease caused by Ebola virus (EBOV). EVD outbreaks typically start from a single case of probable zoonotic transmission, followed by human-to-human transmission via direct contact or contact with infected bodily fluids or contaminated fomites. EVD has a high case–fatality rate; it is characterized by fever, gastrointestinal signs and multiple organ dysfunction syndrome. Diagnosis requires a combination of case definition and laboratory tests, typically real-time reverse transcription PCR to detect viral RNA or rapid diagnostic tests based on immunoassays to detect EBOV antigens. Recent advances in medical countermeasure research resulted in the recent approval of an EBOV-targeted vaccine by European and US regulatory agencies. The results of a randomized clinical trial of investigational therapeutics for EVD demonstrated survival benefits from two monoclonal antibody products targeting the EBOV membrane glycoprotein. New observations emerging from the unprecedented 2013–2016 Western African EVD outbreak (the largest in history) and the ongoing EVD outbreak in the Democratic Republic of the Congo have substantially improved the understanding of EVD and viral persistence in survivors of EVD, resulting in new strategies toward prevention of infection and optimization of clinical management, acute illness outcomes and attendance to the clinical care needs of patients. To date, 12 distinct filoviruses have been described1. outbreak. All FVD outbreaks, with the exception of that The seven filoviruses that have been found in humans caused by TAFV, were characterized by extremely high belong either to the genus Ebolavirus (Bundibugyo virus case–fatality rates (CFRs, also known as lethality). Until (BDBV), Ebola virus (EBOV), Reston virus (RESTV), 2013, the most extensive outbreak, caused by SUDV, Sudan virus (SUDV) and Taï Forest virus (TAFV); Fig. 1) involved 425 cases and 224 deaths (CFR 52.7%)6. The or to the genus Marburgvirus (Marburg virus (MARV) overall limited numbers of FVD cases (1967–2013: and Ravn virus (RAVV))2. The WHO International 2,886 cases including 1,982 deaths4), the typical remote Classification of Diseases Revision 11 (ICD-11) of 2018 and rural locations of outbreaks and the often delayed recognizes two major subcategories of filovirus disease announcement of new outbreaks to the international (FVD): Ebola disease caused by BDBV, EBOV, SUDV community7 have prevented the systematic study of or TAFV, and Marburg disease caused by MARV or clinical FVD in humans. Thus, the commonly used RAVV. Ebola virus disease (EVD) is defined as a disease description of FVD was derived either from observa- only caused by EBOV. This subcategorization of FVD tion of small groups of patients in care settings that were is largely based on the increasing evidence of mole- not well-equipped for diagnosis, treatment and disease cular differences between ebolaviruses and marburg- characterization, or from observations of even smaller viruses, differences that may influence virus–host samples, such as individuals who were transferred from reservoir tropism, pathogenesis and disease phenotype Equatorial Africa to Europe and the USA or who fell sick in accidental primate hosts2. in Europe or the USA after contracting the virus else- Since the discovery of filoviruses in 1967 (REF.3), where. Pathological characterization of FVD via auto- 43 FVD outbreaks (excluding at least five laboratory- psies has been rare7,8. In the absence of extensive human acquired infections) have been recorded in or exported clinical data, FVD could only be defined further via the from Africa4. The epidemiological definition of outbreak use of experimental animal infections9,10. ✉e-mail: kuhnjens@ is one or more cases above the known endemic preva- Until 2013, most EVD outbreaks originated from mail.nih.gov lence. For example, the single case of TAFV infection Middle Africa: the Democratic Republic of the Congo, https://doi.org/10.1038/ recorded in a setting in which FVD had never been Gabon and the Republic of the Congo. From late 2013 s41572-020-0147-3 reported before (Côte d’Ivoire)5 is still considered an to early 2016, EBOV caused the largest outbreak to NATURE REVIEWS | DISEASE PRIMERS | Article citation ID: (2020) 6:13 1 0123456789(); PRIMER 27,28 Author addresses dysfunction . Ultimately, these patients develop robust immune responses leading to clearance of viraemia and a 1 Department of Clinical Sciences, Liverpool School of Tropical Medicine, Liverpool, UK. resolution phase. However, recovery can be complicated 2 Global Health Security Department, Infectious Diseases Institute, Makerere University, by long-lasting clinical sequelae and/or virus persistence Kampala, Uganda. in immune-privileged sites that can lead to disease flares 3Integrated Research Facility at Fort Detrick, Clinical Monitoring Research Program Directorate, Frederick National Laboratory for Cancer Research supported by the and even sexual transmission. In this Primer, we out- National Cancer Institute, Frederick, MD, USA. line the current improved understanding of EVD based 4Department of Medicine, Division of Pulmonary Disease and Critical Care Medicine, on the most recently published human clinical data. Chapel Hill, NC, USA. 5Nebraska Biocontainment Unit, Division of Infectious Diseases, University of Nebraska Epidemiology Medical Center, Omaha, NE, USA. Classic epidemiology 6Microbiology Section, Emory Medical Laboratory, Emory University School of Medicine, Since the discovery of EBOV in 1976 in the Democratic Atlanta, GA, USA. Republic of the Congo (then Zaire)29–31, at least 17 EVD 7 Department of Microbiology, Immunology & Infectious Diseases, Université Laval, outbreaks have originated in Gabon, Guinea, the Quebec City, QC, Canada. Republic of the Congo or Zaire/Democratic Republic of 8Partnership for Ebola Virus Disease Research in Liberia, Monrovia Medical Units ELWA-2 Hospital, Monrovia, Liberia. the Congo. At the time of writing, ~33,604 EBOV infec- 9National Biodefense Analysis and Countermeasures Center, Fort Detrick, Frederick, tions in humans, including 14,742 deaths (average CFR 4,22 MD, USA. 43.8%) are on record , although case numbers differ 10Department of Microbiology and National Emerging Infectious Diseases Laboratories, slightly from source to source. Boston University School of Medicine, Boston, MA, USA. 11Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Outbreaks and transmission. Most outbreaks can be Diseases, National Institutes of Health, Fort Detrick, Frederick, MD, USA. traced back to a single spillover introduction of EBOV into the human population from an unknown reservoir date, which spread from Guinea to other countries in by unknown means. Subsequently, the virus is transmit- Western Africa, leading to 28,652 human infections and ted by direct, typically non-aerosol, human-to-human 11,325 deaths11. The location and scale of the 2013–2016 contact or contact with infected tissues, bodily fluids outbreak was entirely unexpected12. Consequently, local, or contaminated fomites (FIG. 1)4. Based on historical national and international organizations were caught records, EBOV may have been transmitted from its nat- unprepared for an outbreak caused by what, until then, ural reservoir host(s) to humans to cause disease only was considered an exotic pathogen of largely negligi- about 20–30 times (Table 1), although it is probable that ble consequence for global public health13–15. After the limited EVD outbreaks may have been overlooked or not WHO declared the outbreak a Public Health Emergency reported. The potential for infection of an index case and of International Concern, the global and local responses subsequent spread — locally and globally — has been to the outbreak intensified. Ultimately, the outbreak estimated by considering reservoir species distribution, was contained, but it devastated individuals, families, along with governance, communications, isolation, communities, health-care systems and economies16. infrastructure, health care and international connectiv- In most affected countries, the response included the ity32. These predictions are crucial to identify regions that establishment of Ebola (virus disease) Treatment Units require increased surveillance and investments. Tracking (ETUs)17–19, in which medical professionals and biomed- EBOV within the human population after a zoonotic ical scientists managed large cohorts of patients with transmission event can be challenging, especially as the suspected or confirmed EVD in controlled settings. single natural reservoir has not been identified. From this experience, scientists were able to better A strong risk factor linked to human-to-human understand a virus previously best known as a poten- EBOV propagation is contact with infected bodily flu- tial bioweapons agent20,21. In addition to the Western ids33–35. Indeed, infectious EBOV has been recovered African outbreak, an ongoing outbreak in the Ituri, from breast milk, saliva, urine, semen, cerebrospinal Nord-Kivu and Sud-Kivu Provinces of the Democratic fluid, and aqueous humor, in addition to blood and blood Republic of the Congo is the second largest outbreak derivatives, and detected
Load more

Recommended publications
  • A Multicenter, Multi-Outbreak, Randomized, Controlled Safety And
    2018 Ebola MCM RCT Protocol Page 1 of 92 IND#: 125530 CONFIDENTIAL 4 October 2019, version 7.0 A Multicenter, Multi-Outbreak, Randomized, Controlled Safety and Efficacy Study of Investigational Therapeutics for the Treatment of Patients with Ebola Virus Disease Short Title: 2018 Ebola MCM RCT Protocol Sponsored by: Office of Clinical Research Policy and Regulatory Operations (OCRPRO) National Institute of Allergy and Infectious Diseases 5601 Fishers Lane Bethesda, MD 20892 NIH Protocol Number: 19-I-0003 Protocol IND #: 125530 ClinicalTrials.gov Number: NCT03719586 Date: 4 October 2019 Version: 7.0 CONFIDENTIAL This document is confidential. No part of it may be transmitted, reproduced, published, or used by other persons without prior written authorization from the study sponsor and principal investigator. 2018 Ebola MCM RCT Protocol Page 2 of 92 IND#: 125530 CONFIDENTIAL 4 October 2019, version 7.0 KEY ROLES DRC Principal Investigator: Jean-Jacques Muyembe-Tamfum, MD, PhD Director-General, DRC National Institute for Biomedical Research Professor of Microbiology, Kinshasa University Medical School Kinshasa Gombe Democratic Republic of the Congo Phone: +243 898949289 Email: [email protected] Other International Investigators: see Appendix E Statistical Lead: Lori Dodd, PhD Biostatistics Research Branch, DCR, NIAID 5601 Fishers Lane, Room 4C31 Rockville, MD 20852 Phone: 240-669-5247 Email: [email protected] U.S. Principal Investigator: Richard T. Davey, Jr., MD Clinical Research Section, LIR, NIAID, NIH Building 10, Room 4-1479, Bethesda,
    [Show full text]
  • Fighting Ebola: Voices from the Frontline Documenting the Experiences of East African Deployed Experts Who Volunteered to Fight Ebola in West Africa Implemented By
    Fighting Ebola: Voices from the Frontline Documenting the experiences of East African deployed experts who volunteered to fight Ebola in West Africa Implemented by: Fighting Ebola: Voices from the Frontline Documenting the experiences of East African deployed experts who volunteered to fight Ebola in West Africa Contents Introduction . 6 Foreword from the Hon . Jesca Eriyo . 9 Foreword from Dr Zabulon Yoti . .. 10 Foreword from Dr Jackson Amone . 12 Foreword from Dr Irene Lukassowitz . 13 George Acire . 14 Rebecca Racheal Apolot . 17 Sarah Awilo . 22 Dr Mwaniki Collins . 24 Charles Draleku . 26 Emmanuel Ejoku .. 28 Dr Madina Hussein . 31 Loveness Daniel Isojick . 34 Dr Abdulrahman Said Kassim . 36 Teddy Kusemererwa . .. 40 Liliane Luwaga . 43 Dr Appolinaire Manirafasha . 46 Dr Landry Mayigane . 48 James Mugume . 50 Dr Monica Musenero . 52 Doreen Nabawanuka . 55 Dr Bella Nihorimbere . 56 Teresia Wairimu Thuku . 57 Tony Walter Onena . 59 Acknowledgements . 61 4 Fighting Ebola: Voices from the Frontline 5 Introduction ccording to the World Health Organization (WHO), Others, seeing scenes of death and devastation on their The East African Community Secretariat, in collaboration the Ebola epidemic that occurred in West Africa television screens, just felt compelled to do whatever they with the Federal Government of Germany through the Abetween 2014 and 2016 killed over 11,000 people could to help . Given that so many West African health GIZ-coordinated Support to Pandemic Preparedness in out of the almost 30,000 that were infected . From one
    [Show full text]
  • COVID-19) Pandemic on National Antimicrobial Consumption in Jordan
    antibiotics Article An Assessment of the Impact of Coronavirus Disease (COVID-19) Pandemic on National Antimicrobial Consumption in Jordan Sayer Al-Azzam 1, Nizar Mahmoud Mhaidat 1, Hayaa A. Banat 2, Mohammad Alfaour 2, Dana Samih Ahmad 2, Arno Muller 3, Adi Al-Nuseirat 4 , Elizabeth A. Lattyak 5, Barbara R. Conway 6,7 and Mamoon A. Aldeyab 6,* 1 Clinical Pharmacy Department, Jordan University of Science and Technology, Irbid 22110, Jordan; [email protected] (S.A.-A.); [email protected] (N.M.M.) 2 Jordan Food and Drug Administration (JFDA), Amman 11181, Jordan; [email protected] (H.A.B.); [email protected] (M.A.); [email protected] (D.S.A.) 3 Antimicrobial Resistance Division, World Health Organization, Avenue Appia 20, 1211 Geneva, Switzerland; [email protected] 4 World Health Organization Regional Office for the Eastern Mediterranean, Cairo 11371, Egypt; [email protected] 5 Scientific Computing Associates Corp., River Forest, IL 60305, USA; [email protected] 6 Department of Pharmacy, School of Applied Sciences, University of Huddersfield, Huddersfield HD1 3DH, UK; [email protected] 7 Institute of Skin Integrity and Infection Prevention, University of Huddersfield, Huddersfield HD1 3DH, UK * Correspondence: [email protected] Citation: Al-Azzam, S.; Mhaidat, N.M.; Banat, H.A.; Alfaour, M.; Abstract: Coronavirus disease 2019 (COVID-19) has overlapping clinical characteristics with bacterial Ahmad, D.S.; Muller, A.; Al-Nuseirat, respiratory tract infection, leading to the prescription of potentially unnecessary antibiotics. This A.; Lattyak, E.A.; Conway, B.R.; study aimed at measuring changes and patterns of national antimicrobial use for one year preceding Aldeyab, M.A.
    [Show full text]
  • Ebola Virus Disease
    Outbreaks Chronology: Ebola Virus Disease Known Cases and Outbreaks of Ebola Virus Disease, in Reverse Chronological Order: Reported number (%) of Reported deaths Ebola number of among Year(s) Country subtype human cases cases Situation August- Democratic Ebola virus 66 49 (74%) Outbreak occurred in November 2014 Republic of multiple villages in the Congo the Democratic Republic of the Congo. The outbreak was unrelated to the outbreak of Ebola in West Africa. March 2014- Multiple Ebola virus 28652 11325 Outbreak across Present countries multiple countries in West Africa. Number of patients is constantly evolving due to the ongoing investigation. 32 November 2012- Uganda Sudan virus 6* 3* (50%) Outbreak occurred in January 2013 the Luwero District. CDC assisted the Ministry of Health in the epidemiologic and diagnostic aspects of the outbreak. Testing of samples by CDC's Viral Special Pathogens Branch occurred at UVRI in Entebbe. 31 June-November Democratic Bundibugyo 36* 13* (36.1%) Outbreak occurred in 2012 Republic of virus DRC’s Province the Congo Orientale. Laboratory support was provided through CDC and the Public Health Agency of Canada (PHAC)’s field laboratory in Isiro, as well as through the CDC/UVRI lab in Uganda. The outbreak in DRC had no epidemiologic link to the near contemporaneous Ebola outbreak in the Kibaale district of Uganda. 31 June-October Uganda Sudan virus 11* 4* (36.4%) Outbreak occurred in 2012 the Kibaale District of Uganda. Laboratory tests of blood samples were conducted by the UVRI and the CDC. 31 May 2011 Uganda Sudan virus 1 1 (100%) The Uganda Ministry of Health informed the public a patient with suspected Ebola Hemorrhagic fever died on May 6, 2011 in the Luwero district, Uganda.
    [Show full text]
  • Targeting Ebola 2015
    Agenda of Targeting Ebola 2015 May 28-29, 2015 Institut Pasteur - Paris, France www.targeting-ebola.com Welcome Note On behalf of the Targeting Infectious Diseases Committee, the Institut Pasteur, the COPED and the Task Force for Ebola in France, it is our pleasure to announce the organization of the International Congress on Targeting Ebola 2015 which will be held on May 28-29, 2015 at Pasteur Institute, Paris, France. The World Health Organization (WHO) was notified on March 23, 2014, of an outbreak of EVD in Guinea. The disease soon spread to the bordering countries of Liberia and Sierra Leone, which are the most severely affected countries. On August 8, 2014, the epidemic was declared a “public health emergency of international concern” (WHO Ebola Response Team, NEJM 2014, 371, 1481). Suspected cases of EVD have since been reported in seven affected countries (Guinea, Liberia, Nigeria, Senegal, Sierra Leone, Spain, and the United States of America). Unprecedented in scale and geographical distribution since the identification of Ebola in 1976, the current epidemic has an apparent overall case-fatality ratio of about 70%; but it is suspected that many more cases have gone unrecorded. On May 2015, more than 10.000 deaths and 30.000 cases had been reported in Sierra Leone, Liberia and Guinea, according to the WHO. Globally, the situation is improving. The epidemic is over in Liberia, however in Guinea and Sierra Leone, the initial decline observed a few weeks ago is stable with approximately 20 new cases / week. The vast majority of cases are reported from the western prefecture of Forecariah, which borders the Sierra Leonean district of Kambia.
    [Show full text]
  • Clinical Trial Protocol Sponsored by OCRPRO
    PREVAIL IV: Double-blind, Randomized, Two-phase, Placebo-controlled, Phase II Trial of GS-5734 to Assess the Antiviral Activity, Longer-term Clearance of Ebola Virus, and Safety in Male Ebola Survivors with Evidence of Ebola Virus Persistence in Semen NIAID Protocol Number: 16-I-N137 Version Number: 10.0 Date: May 13, 2019 Investigational New Drug (IND) Number: 130621 IND Sponsor: Office of Clinical Research Policy and Regulatory Operations (OCRPRO), Division of Clinical Research (DCR), National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH) Local Liberian Medical Officer: Ian Wachekwa, MD Safety Medical Officer John F. Kennedy Hospital 21 Street, Sinkor Monrovia, Liberia [email protected] (t) +231880890907 Sponsor Medical Monitor: Alan Lifson, MD, MPH Professor, Division of Epidemiology and Community Health School of Public Health University of Minnesota 1300 S. Second St., Ste. 300 Minneapolis, MN 55454 (t) 612-626-9697 (f) 612-624-0315 [email protected] Pharmaceutical Support Provided by: Gilead Sciences, Inc. PREVAIL IV Version 10.0 13 May 2019 Conducted by: Liberia-US Joint Clinical Research Partnership also known as the Partnership for Research on Ebola Virus in Liberia (PREVAIL) Page 2 of 77 PREVAIL IV Version 10.0 13 May 2019 Key Roles NIH Principal Investigator: Elizabeth S. Higgs, MD, MIA, DTMH Division of Clinical Research, NIAID [email protected] (c) +301-768-3947 skype: libby.higgs2 Liberian Principal Investigator: Dehkontee Gayedyu-Dennis, MD PREVAIL [email protected] (t) +231-886-538-810
    [Show full text]
  • Chapter 1 BEFORE EBOLA
    10/12/2017 Preparing for a Crisis: Lessons Learned at Emory Before, During, and After Ebola Jay B. Varkey, MD On behalf of the Chapter 1 BEFORE EBOLA 2 1 10/12/2017 The Story of Emile Meliandou, Guinea, December 2013 Photograph by Suzanne Beukes, UNICEF The Story of Emile Meliandou, Guinea, December 2013 AM Saéz et al, EMBO Mole Med 2015 2 10/12/2017 The Story of Emile Meliandou, Guinea, December 2013 The Story of Emile Meliandou, Guinea, December 2013 AM Saéz et al, EMBO Mole Med 2015 3 10/12/2017 The Story of Emile Meliandou, Guinea, December 2013 Photograph by Suzanne Beukes, UNICEF Spread of a “Cholera‐Like Illness” MeliandouConakry, 12/2013 – 3/2014 S Baize, N Engl J Med 2014 4 10/12/2017 Ebola Virus Disease (EVD) Spread to Liberia and Sierra Leone 4/2014 ELWA Ebola Treatment Unit (ETU) Monrovia, Liberia 6/2014 – 7/2014 Photograph by Bethany Fankhauser, SIM 5 10/12/2017 ELWA ETU Monrovia, Liberia 6/2014 – 7/2014 Photograph by Bethany Fankhauser, SIM ELWA ETU Monrovia, Liberia July 2014 • 40 patients with EVD treated • 39 of 40 died • July 22‐23: 2 American humanitarians working in the ELWA ETC develop a fever…. • Initially treated for malaria • Laboratory testing confirmed Ebola Virus Disease 6 10/12/2017 Ebola Virus Disease and Emory July 30, 2014 • Emory University Hospital was asked to receive the first patients with confirmed Ebola virus disease to be treated in the United States • 2 American humanitarians had become infected while working in an ETU in Monrovia, Liberia • Patient 1: 33 yo male physician • Patient 2: 59 yo female medical missionary • Expected arrival time unclear but Emory was asked to be ready to receive the 1st patient within 72 hours.
    [Show full text]
  • Replication of Marburg Virus in Human Endothelial Cells a Possible Mechanism for the Development of Viral Hemorrhagic Disease
    Replication of Marburg Virus in Human Endothelial Cells A Possible Mechanism for the Development of Viral Hemorrhagic Disease Hans-Joachim Schnittler,$ Friederike Mahner, * Detlev Drenckhahn, * Hans-Dieter Klenk, * and Heinz Feldmann * *Institut fir Virologie, Philipps-Universitdt Marburg, 3550 Marburg, Germany; tInstitutftirAnatomie, Universitdt Wiirzburg, 8700 Wiirzburg, Germany Abstract Rhabdoviridae, within the new proposed order Mononegavi- Marburg and Ebola virus, members of the family Filoviridae, rales (10). Virions are composed of a helical nucleocapsid cause a severe hemorrhagic disease in humans and primates. surrounded by a lipid envelope. The genome is nonsegmented, The disease is characterized as a pantropic virus infection often of negative sense, and 19 kb in length (3, 11, 12). Virion parti- resulting in a fulminating shock associated with hemorrhage, cles contain at least seven structural proteins (8, 13-16). and death. All known histological and pathophysiological pa- Filovirus infections have several pathological features in rameters of the disease are not sufficient to explain the devas- common with other severe viral hemorrhagic fevers such as tating symptoms. Previous studies suggested a nonspecific de- Lassa fever, hemorrhagic fever with renal syndrome, and struction of the endothelium as a possible mechanism. Con- Dengue hemorrhagic fever (5). Among these viruses, filovi- cerning the important regulatory functions of the endothelium ruses cause the highest case-fatality rates ( - 35% for MBG [61 (blood pressure, antithrombogenicity, homeostasis), we exam- and up to 90% for EBO, subtype Zaire [17]) and the most ined Marburg virus replication in primary cultures of human severe hemorrhagic manifestations. The pathophysiologic endothelial cells and organ cultures of human umbilical cord events that make filovirus infections of humans so devastating veins.
    [Show full text]
  • Rabbit Anti-Marburgvirus (MARV) VLP Pab ELISA Data
    4 Research Court, Suite 300 Rockville, MD 20850 877-411-2041 [email protected] Rabbit anti-Marburgvirus (MARV) VLP pAb ELISA Data: Catalog #: 04-0005 IgG IgG + IgG + Lot #: MMIG201001IBT Dilution MMARV ZEBOV 1:X Antigen Antigen Immunogen: MARV (Musoke strain) Virus-like Particles (VLPs) containing glycoprotein (GP) 1000 3.30 2.66 Nucleoprotein (NP), and viral protein (VP40). 3162 3.07 1.80 10000 2.68 0.89 Description: Protein A purified rabbit polyclonal 31623 1.87 0.37 antibody reactive to MARV VLP raised in New 100000 0.99 0.14 Zealand white rabbits. 316228 0.43 0.05 1000000 0.17 0.02 Supplied: 0.5 mg of antibody is supplied in PBS at a concentration of 5.75 mg/mL. 0.01% Sodium azide has been added. -Antigen is coated on ELISA plates overnight. -Add 200µl blocking buffer then wash wells with Clonality: Polyclonal PBST. -Antiserum is diluted semi-log. Relevance: the filovirus Marburgvirus is a -Incubate antibody for 2 hour. Category A (NIAID) and HHS select agent. -Wash unbound antibodies and add HRP- Recommended Dilutions: conjugated anti-rabbit IgG. -Wash plates and add substrate to develop color for ELISA: Assay-dependent dilution. 20 minutes. WB: Assay-dependent dilution -Read absorbance at 650nm. Amount of color is directly proportional to amount of antibodies. Storage: 2-3 weeks +4oC, -20◦C long term Western Blot Cross Reactivity: Historical data showed some cross-reactivity with Ebola Virus (EBOV) and -Antiserum recognizes Marburg musoke Sudan Virus (SUDV) VLP’s, most likely due to glycoprotein, nucleoprotein, and VP40 antibodies against Baculovirus proteins since the VLP’s were expressed in SF9-Baculovirus system.
    [Show full text]
  • Comparison and Evaluation of Real-Time Taqman PCR for Detection and Quantification of Ebolavirus
    viruses Article Comparison and Evaluation of Real-Time Taqman PCR for Detection and Quantification of Ebolavirus Yi Huang 1,* , Shuqi Xiao 2 and Zhiming Yuan 1,* 1 National Biosafety Laboratory, Chinese Academy of Sciences, Wuhan 430020, China 2 Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430020, China; [email protected] * Correspondence: [email protected] (Y.H.); [email protected] (Z.Y.) Abstract: Given that ebolavirus causes severe and frequently lethal disease, its rapid and accurate detection using available and validated methods is essential for controlling infection. Real-time reverse-transcription PCR (RT-PCR) has proven to be an invaluable tool for ebolaviruses diagnostics. Many assays with different targets have been developed, but they have not been externally compared or validated, and limits of detection are not uniformly reported. Here we compared and evaluated the sensitivity, reproducibility and specificity of 23 in-house assays under the same conditions. Our results showed that these assays were highly gene- and species- specific when evaluated using in vitro RNA transcripts and viral RNA, and the potential limits of detection were uniformly reported 2 6 ranging from 10 to 10 in vitro synthesized RNA transcripts copies perµL and 1–100 TCID50/mL. The comparison of these assays indicated that those targeting the more conservative NP gene could be the better option for EVD case definition and quantitative measurement because of its higher sensitivity for the same species. Our analysis could contribute to the standardization of ebolavirus detection and quantification assays, which can offer a better understanding of the meaning of results across laboratories and time points, as well as make them easy to implement, especially under outbreak conditions.
    [Show full text]
  • Kinase Inhibitors and Nucleoside Analogues As Novel Therapies to Inhibit HIV-1 Or ZEBOV Replication
    Kinase Inhibitors and Nucleoside Analogues as Novel Therapies to Inhibit HIV-1 or ZEBOV Replication by Stephen D. S. McCarthy A thesis submitted in conformity with the requirements for the degree of Doctor of Philosophy Department of Laboratory Medicine and Pathobiology University of Toronto © Copyright by Stephen D. S. McCarthy (2017) Kinase Inhibitors and Nucleoside Analogues as Novel Therapies to Inhibit HIV-1 or ZEBOV Replication Stephen D.S. McCarthy Doctor of Philosophy Graduate Department of Laboratory Medicine and Pathobiology University of Toronto 2017 Abstract Without a vaccine for Human Immunodeficiency Virus type 1 (HIV-1), or approved therapy for treating Zaire Ebolavirus (ZEBOV) infection, new means to treat either virus during acute infection are under intense investigation. Repurposing tyrosine kinase inhibitors of known specificity may not only inhibit HIV-1 replication, but also treat associated inflammation or neurocognitive disorders caused by chronic HIV-1 infection. Moreover, tyrosine kinase inhibitors may effectively treat other infections, including ZEBOV. In addition, established nucleoside/nucleotide analogues that effectively inhibit HIV-1 infection, could also be repurposed to inhibit ZEBOV replication. In this work the role of two host cell kinases, cellular protoncogene SRC (c-SRC) and Protein Tyrosine Kinase 2 Beta (PTK2B), were found to have key roles during early HIV-1 replication in primary activated CD4+ T-cells ex vivo. siRNA knockdown of either kinase increased intracellular reverse transcripts and decreased nuclear proviral integration, suggesting they act at the level of pre-integration complex (PIC) formation or PIC nuclear translocation. c-SRC siRNA knockdown consistently reduced p24 levels of IIIB(X4) and Ba-L(R5) infection, or luciferase ii activity of HXB2(X4) or JR-FL(R5) recombinant viruses, prompting further drug inhibition studies of this kinase.
    [Show full text]
  • Accelerating R&D for Emerging Infectious Diseases: Lessons
    Accelerating R&D for Emerging Infectious Diseases: Lessons Learnt From Ebola Chao Li China CDC Jingyi Chen Harvard University Jiyan Ma Peking University Health Science Centre Yangmu Huang ( [email protected] ) Peking University Health Science Centre https://orcid.org/0000-0002-3660-1276 Research Article Keywords: Ebola Virus Disease, Emerging Infectious Disease, Research and Development Posted Date: July 2nd, 2020 DOI: https://doi.org/10.21203/rs.3.rs-38612/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/12 Abstract Objective: The R&D explosion for Ebola virus disease (EVD) during the 2014-2016 outbreak led to the successful development of high-quality vaccines performed by China and the U.S. This study aims to compare the R&D activities of Ebola-related medical products in two countries, as a way to present the inuential factors of R&D for emerging infectious disease (EID) and to provide suggestions for timely and ecient R&D response to the COVID-19 pandemic. Methods: In this comparative study, R&D activities were analyzed in terms of research funding, scientic research outputs, R&D timeline, and government incentive and coordinated mechanisms. Quantitative analysis was performed using data retrieved from national websites, clinical trial registries and databases.Qualitative semi-structured interviews were conducted to explore perspectives of fteen key informants from EID eld, especially of those involved in Ebola product development. Findings: The funding gap between China and the U.S. was signicant before 2014 and narrowed after the Ebola outbreak. Both research teams started basic studies prior to the Ebola outbreak; however, the U.S.
    [Show full text]