Simultaneous Detection of Ebola Virus and Pathogens Associated With Hemorrhagic Fever by an Oligonucleotide Microarray

Ebola virus infection causes severe hemorrhagic fever, and its mortality rates varied from 25 to 90% in the previous outbreaks. The highly infectious and lethal nature of this virus highlights the need for reliable and sensitive diagnostic methods to distinguish it from other diseases present with similar clinical symptoms. Based on multiplex polymerase chain reaction (PCR) and oligonucleotide microarray technology, a cost-effective, multipathogen and high-throughput method was developed for simultaneous detection of Ebola virus and other pathogens associated with hemorrhagic fever, including Marburg virus, Lassa fever virus, Junin virus, Machupo virus, Rift Valley fever virus, Crimean-Congo hemorrhagic fever virus, malaria parasite, hantavirus, severe fever with thrombocytopenia syndrome virus, dengue virus, yellow fever virus, Chikungunya virus, influenza A virus, and influenza B virus. This assay had an excellent specificity for target pathogens, without overlap signal between the probes. The limit of detection was approximately 103 pathogen copies/μl. A total of 60 positive nucleic acid samples for different pathogens were detected, a concordance of 100% was observed between microarray assay and real-time PCR analysis. Consequently, the described oligonucleotide microarray may be specific and sensitive assay for diagnosis and surveillance of infections caused by Ebola virus and other species of hemorrhagic fever pathogens.

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Chikungunya and O’nyong-nyong Viruses in Uganda: Implications for Diagnostics

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz001 ◽

2019 ◽

Vol 6 (3) ◽

Cited By ~ 3

Author(s):

Tamara L Clements ◽

Cynthia A Rossi ◽

Amanda K Irish ◽

Hannah Kibuuka ◽

Leigh Anne Eller ◽

...

Keyword(s):

Ebola Virus ◽

Yellow Fever Virus ◽

Rift Valley Fever Virus ◽

Hemorrhagic Fever ◽

Marburg Virus ◽

Fever Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Lassa Virus ◽

Antibody Prevalence ◽

Valley Fever

Abstract Background A serosurvey of healthy blood donors provided evidence of hemorrhagic fever and arthropod-borne virus infections in Uganda. Methods Antibody prevalence to arthropod-borne and hemorrhagic fever viruses in human sera was determined using enzyme-linked immunosorbent assay (ELISA) and plaque reduction neutralization test (PRNT). Results The greatest antibody prevalence determined by ELISA was to chikungunya virus (CHIKV) followed in descending order by West Nile virus (WNV), Crimean-Congo hemorrhagic fever virus (CCHFV), Ebola virus (EBOV), dengue virus (DEN), yellow fever virus (YFV), Rift Valley fever virus (RVFV), Marburg virus (MARV), and Lassa virus (LASV). Further investigation of CHIKV-positive sera demonstrated that the majority of antibody responses may likely be the result of exposure to the closely related alphavirus o’nyong-nyong virus (ONNV). Conclusions As the use of highly specific and sensitive polymerase chain reaction–based assays becomes the diagnostic standard without the corresponding use of the less sensitive but more broadly reactive immunological-based assays, emerging and re-emerging outbreaks will be initially missed, illustrating the need for an orthogonal system for the detection and identification of viruses causing disease.

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Yellow Fever Virus: Diagnostics for a Persistent Arboviral Threat

Journal of Clinical Microbiology ◽

10.1128/jcm.00827-18 ◽

2018 ◽

Vol 56 (10) ◽

Cited By ~ 9

Author(s):

Jesse J. Waggoner ◽

Alejandra Rojas ◽

Benjamin A. Pinsky

Keyword(s):

Yellow Fever ◽

Yellow Fever Virus ◽

Current Knowledge ◽

Hemorrhagic Fever ◽

Clinical Diagnostics ◽

Fever Virus ◽

Diagnostic Methods ◽

Effective Vaccine ◽

Molecular Testing ◽

Viral Culture

ABSTRACTYellow fever (YF) is the prototypical hemorrhagic fever and results from infection with yellow fever virus (YFV), which is endemic to regions of Africa and South America. Despite the availability of an effective vaccine, YFV continues to cause disease throughout regions where it is endemic, including intermittent large outbreaks among undervaccinated populations. A number of diagnostic methods and assays have been described for the detection of YFV infection, including viral culture, molecular testing, serology, and antigen detection. Commercial diagnostics are not widely available, and testing is generally performed at a small number of reference laboratories. The goal of this article, therefore, is to review available clinical diagnostics for YFV, which may not be familiar to many practitioners outside areas where it is endemic. Additionally, we identify gaps in our current knowledge about YF that pertain to diagnosis and describe interventions that may improve YFV detection.

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Yellow Fever Outbreak in Eastern Senegal, 2020–2021

Viruses ◽

10.3390/v13081475 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1475

Author(s):

Moussa Moïse Diagne ◽

Marie Henriette Dior Ndione ◽

Alioune Gaye ◽

Mamadou Aliou Barry ◽

Diawo Diallo ◽

...

Keyword(s):

Yellow Fever ◽

Yellow Fever Virus ◽

Mosquito Species ◽

Virus Transmission ◽

Genomic Analysis ◽

Hemorrhagic Fever ◽

Fever Virus ◽

World Health ◽

Effective Vaccine ◽

Ministry Of Health

Yellow fever virus remains a major threat in low resource countries in South America and Africa despite the existence of an effective vaccine. In Senegal and particularly in the eastern part of the country, periodic sylvatic circulation has been demonstrated with varying degrees of impact on populations in perpetual renewal. We report an outbreak that occurred from October 2020 to February 2021 in eastern Senegal, notified and managed through the synergistic effort yellow fever national surveillance implemented by the Senegalese Ministry of Health in collaboration with the World Health Organization, the countrywide 4S network set up by the Ministry of Health, the Institut Pasteur de Dakar, and the surveillance of arboviruses and hemorrhagic fever viruses in human and vector populations implemented since mid 2020 in eastern Senegal. Virological analyses highlighted the implication of sylvatic mosquito species in virus transmission. Genomic analysis showed a close relationship between the circulating strain in eastern Senegal, 2020, and another one from the West African lineage previously detected and sequenced two years ago from an unvaccinated Dutch traveler who visited the Gambia and Senegal before developing signs after returning to Europe. Moreover, genome analysis identified a 6-nucleotide deletion in the variable domain of the 3′UTR with potential impact on the biology of the viral strain that merits further investigations. Integrated surveillance of yellow fever virus but also of other arboviruses of public health interest is crucial in an ecosystem such as eastern Senegal.

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A point-of-care diagnostic for differentiating Ebola from endemic febrile diseases

Science Translational Medicine ◽

10.1126/scitranslmed.aat0944 ◽

2018 ◽

Vol 10 (471) ◽

pp. eaat0944 ◽

Cited By ~ 19

Author(s):

David Sebba ◽

Alexander G. Lastovich ◽

Melody Kuroda ◽

Eric Fallows ◽

Joshua Johnson ◽

...

Keyword(s):

Ebola Virus ◽

Clinical Symptoms ◽

Point Of Care ◽

Hemorrhagic Fever ◽

Diagnostic Methods ◽

Outbreak Detection ◽

Molecular Diagnostic ◽

Clinical Samples ◽

Live Virus ◽

And Control

Hemorrhagic fever outbreaks such as Ebola are difficult to detect and control because of the lack of low-cost, easily deployable diagnostics and because initial clinical symptoms mimic other endemic diseases such as malaria. Current molecular diagnostic methods such as polymerase chain reaction require trained personnel and laboratory infrastructure, hindering diagnostics at the point of need. Although rapid tests such as lateral flow can be broadly deployed, they are typically not well-suited for differentiating among multiple diseases presenting with similar symptoms. Early detection and control of Ebola outbreaks require simple, easy-to-use assays that can detect and differentiate infection with Ebola virus from other more common febrile diseases. Here, we developed and tested an immunoassay technology that uses surface-enhanced Raman scattering (SERS) tags to simultaneously detect antigens from Ebola, Lassa, and malaria within a single blood sample. Results are provided in <30 min for individual or batched samples. Using 190 clinical samples collected from the 2014 West African Ebola outbreak, along with 163 malaria positives and 233 negative controls, we demonstrated Ebola detection with 90.0% sensitivity and 97.9% specificity and malaria detection with 100.0% sensitivity and 99.6% specificity. These results, along with corresponding live virus and nonhuman primate testing of an Ebola, Lassa, and malaria 3-plex assay, indicate the potential of the SERS technology as an important tool for outbreak detection and clinical triage in low-resource settings.

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Simultaneous detection of Dengue virus, Chikungunya virus, Zika virus, Yellow fever virus and West Nile virus

Journal of Virological Methods ◽

10.1016/j.jviromet.2019.03.014 ◽

2019 ◽

Vol 268 ◽

pp. 53-55 ◽

Cited By ~ 3

Author(s):

José A. Boga ◽

Marta E. Alvarez-Arguelles ◽

Susana Rojo-Alba ◽

Mercedes Rodríguez ◽

María de Oña ◽

...

Keyword(s):

West Nile Virus ◽

Dengue Virus ◽

Yellow Fever ◽

Zika Virus ◽

Chikungunya Virus ◽

Yellow Fever Virus ◽

Simultaneous Detection ◽

Fever Virus ◽

West Nile ◽

Virus Zika

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Macaque Monoclonal Antibodies Targeting Novel Conserved Epitopes within Filovirus Glycoprotein

Journal of Virology ◽

10.1128/jvi.02172-15 ◽

2015 ◽

Vol 90 (1) ◽

pp. 279-291 ◽

Cited By ~ 48

Author(s):

Zhen-Yong Keck ◽

Sven G. Enterlein ◽

Katie A. Howell ◽

Hong Vu ◽

Sergey Shulenin ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Nonhuman Primates ◽

Ebola Virus ◽

Virus Disease ◽

Protective Efficacy ◽

Hemorrhagic Fever ◽

Marburg Virus ◽

Human Pathogens ◽

Content Type ◽

The Core

ABSTRACTFiloviruses cause highly lethal viral hemorrhagic fever in humans and nonhuman primates. Current immunotherapeutic options for filoviruses are mostly specific to Ebola virus (EBOV), although other members ofFiloviridaesuch as Sudan virus (SUDV), Bundibugyo virus (BDBV), and Marburg virus (MARV) have also caused sizeable human outbreaks. Here we report a set of pan-ebolavirus and pan-filovirus monoclonal antibodies (MAbs) derived from cynomolgus macaques immunized repeatedly with a mixture of engineered glycoproteins (GPs) and virus-like particles (VLPs) for three different filovirus species. The antibodies recognize novel neutralizing and nonneutralizing epitopes on the filovirus glycoprotein, including conserved conformational epitopes within the core regions of the GP1 subunit and a novel linear epitope within the glycan cap. We further report the first filovirus antibody binding to a highly conserved epitope within the fusion loop of ebolavirus and marburgvirus species. One of the antibodies binding to the core GP1 region of all ebolavirus species and with lower affinity to MARV GP cross neutralized both SUDV and EBOV, the most divergent ebolavirus species. In a mouse model of EBOV infection, this antibody provided 100% protection when administered in two doses and partial, but significant, protection when given once at the peak of viremia 3 days postinfection. Furthermore, we describe novel cocktails of antibodies with enhanced protective efficacy compared to individual MAbs. In summary, the present work describes multiple novel, cross-reactive filovirus epitopes and innovative combination concepts that challenge the current therapeutic models.IMPORTANCEFiloviruses are among the most deadly human pathogens. The 2014-2015 outbreak of Ebola virus disease (EVD) led to more than 27,000 cases and 11,000 fatalities. While there are five species ofEbolavirusand several strains of marburgvirus, the current immunotherapeutics primarily target Ebola virus. Since the nature of future outbreaks cannot be predicted, there is an urgent need for therapeutics with broad protective efficacy against multiple filoviruses. Here we describe a set of monoclonal antibodies cross-reactive with multiple filovirus species. These antibodies target novel conserved epitopes within the envelope glycoprotein and exhibit protective efficacy in mice. We further present novel concepts for combination of cross-reactive antibodies against multiple epitopes that show enhanced efficacy compared to monotherapy and provide complete protection in mice. These findings set the stage for further evaluation of these antibodies in nonhuman primates and development of effective pan-filovirus immunotherapeutics for use in future outbreaks.

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Multicenter Evaluation of the ePlex Respiratory Pathogen Panel for the Detection of Viral and Bacterial Respiratory Tract Pathogens in Nasopharyngeal Swabs

Journal of Clinical Microbiology ◽

10.1128/jcm.01658-17 ◽

2017 ◽

Vol 56 (2) ◽

Cited By ~ 21

Author(s):

N. Esther Babady ◽

Matthew R. England ◽

Kristen L. Jurcic Smith ◽

Taojun He ◽

Dona Saumya Wijetunge ◽

...

Keyword(s):

Influenza A ◽

Simultaneous Detection ◽

Respiratory Pathogen ◽

Influenza B Virus ◽

Nasopharyngeal Swab ◽

Influenza B ◽

Content Type ◽

Subtype B ◽

Percent Agreement ◽

Virus Influenza

ABSTRACT The performance of the new ePlex Respiratory Pathogen (RP) panel (GenMark Diagnostics) for the simultaneous detection of 19 viruses (influenza A virus; influenza A H1 virus; influenza A 2009 H1 virus; influenza A H3 virus; influenza B virus; adenovirus; coronaviruses [HKU1, OC43, NL63, and 229E]; human rhinovirus/enterovirus; human metapneumovirus; parainfluenza viruses 1, 2, 3, and 4; and respiratory syncytial virus [RSV] [RSV subtype A and RSV subtype B]) and 2 bacteria (Mycoplasma pneumoniae and Chlamydia pneumoniae) was evaluated. Prospectively and retrospectively collected nasopharyngeal swab (NPS) specimens (n = 2,908) were evaluated by using the ePlex RP panel, with the bioMérieux/BioFire FilmArray Respiratory Panel (BioFire RP) as the comparator method. Discordance analysis was performed by using target-specific PCRs and bidirectional sequencing. The reproducibility of the assay was evaluated by using reproducibility panels comprised of 6 pathogens. The overall agreement between the ePlex RP and BioFire RP results was >95% for all targets. Positive percent agreement with the BioFire RP result for viruses ranged from 85.1% (95% confidence interval [CI], 80.2% to 88.9%) to 95.1% (95% CI, 89.0% to 97.9%), while negative percent agreement values ranged from 99.5% (95% CI, 99.1% to 99.7%) to 99.8% (95% CI, 99.5% to 99.9%). Additional testing of discordant targets (12%; 349/2,908) confirmed the results of ePlex RP for 38% (131/349) of samples tested. Reproducibility was 100% for all targets tested, with the exception of adenovirus, for which reproducibilities were 91.6% at low virus concentrations and 100% at moderate virus concentrations. The ePlex RP panel offers a new, rapid, and sensitive “sample-to-answer” multiplex panel for the detection of the most common viral and bacterial respiratory pathogens.

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Mapping of Genomic Segments of Influenza B Virus Strains by an Oligonucleotide Microarray Method

Journal of Clinical Microbiology ◽

10.1128/jcm.42.12.5793-5801.2004 ◽

2004 ◽

Vol 42 (12) ◽

pp. 5793-5801 ◽

Cited By ~ 16

Author(s):

A. V. Ivshina ◽

G. M. Vodeiko ◽

V. A. Kuznetsov ◽

D. Volokhov ◽

R. Taffs ◽

...

Keyword(s):

Oligonucleotide Microarray ◽

Influenza B Virus ◽

Influenza B ◽

B Virus ◽

Virus Strains

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Mosquito-Borne Viral Pathogens Detected in Zambia: A Systematic Review

Pathogens ◽

10.3390/pathogens10081007 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1007

Author(s):

Rachel Milomba Velu ◽

Geoffrey Kwenda ◽

Liyali Libonda ◽

Caroline Cleopatra Chisenga ◽

Bumbangi Nsoni Flavien ◽

...

Keyword(s):

Systematic Review ◽

Viral Vectors ◽

Yellow Fever Virus ◽

Rift Valley Fever Virus ◽

Fever Virus ◽

Diagnostic Methods ◽

Control Measures ◽

Viral Pathogens ◽

Valley Fever ◽

Effective Prevention

Emerging and re-emerging mosquito-borne viral diseases are a threat to global health. This systematic review aimed to investigate the available evidence of mosquito-borne viral pathogens reported in Zambia. A search of literature was conducted in PubMed and Google Scholar for articles published from 1 January 1930 to 30 June 2020 using a combination of keywords. Eight mosquito-borne viruses belonging to three families, Togaviridae, Flaviviridae and Phenuiviridae were reported. Three viruses (Chikungunya virus, Mayaro virus, Mwinilunga virus) were reported among the togaviruses whilst four (dengue virus, West Nile virus, yellow fever virus, Zika virus) were among the flavivirus and only one virus, Rift Valley fever virus, was reported in the Phenuiviridae family. The majority of these mosquito-borne viruses were reported in Western and North-Western provinces. Aedes and Culex species were the main mosquito-borne viral vectors reported. Farming, fishing, movement of people and rain patterns were among factors associated with mosquito-borne viral infection in Zambia. Better diagnostic methods, such as the use of molecular tools, to detect the viruses in potential vectors, humans, and animals, including the recognition of arboviral risk zones and how the viruses circulate, are important for improved surveillance and design of effective prevention and control measures.

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Ebola and Marburg Viruses

Journal of Disaster Research ◽

10.20965/jdr.2011.p0381 ◽

2011 ◽

Vol 6 (4) ◽

pp. 381-389 ◽

Cited By ~ 7

Author(s):

Eri Nakayama ◽

◽

Ayato Takada

Keyword(s):

Nonhuman Primates ◽

Ebola Virus ◽

Hemorrhagic Fever ◽

Disease Outbreak ◽

Marburg Virus ◽

Virus Group ◽

Imported Cases ◽

Control Disease ◽

Animal Reservoirs ◽

Biosafety Level

Ebola and Marburg viruses, members of the filovirus family, cause severe hemorrhagic fever in human and nonhuman primates and are classified as biosafety level 4 agents. No effective filovirus-specific prophylaxis or treatment is yet commercially available. Filovirus species vary genetically, with one in the Marburg virus group and five in the Ebola virus group. Epidemiological efforts to prevent outbreaks lie mainly in identifying natural animal reservoirs. Increasingly frequent outbreaks in Africa and concerns about bioterrorism and imported cases in nonendemic areas point to the importance of public health in two ways – finding strategies to control disease outbreak and developing effective vaccines and drugs.

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