Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Mar 17;9(3):e0003631.
doi: 10.1371/journal.pntd.0003631. eCollection 2015 Mar.

Discovery of Novel Rhabdoviruses in the Blood of Healthy Individuals From West Africa

Free PMC article

Discovery of Novel Rhabdoviruses in the Blood of Healthy Individuals From West Africa

Matthew H Stremlau et al. PLoS Negl Trop Dis. .
Free PMC article


Next-generation sequencing (NGS) has the potential to transform the discovery of viruses causing unexplained acute febrile illness (UAFI) because it does not depend on culturing the pathogen or a priori knowledge of the pathogen's nucleic acid sequence. More generally, it has the potential to elucidate the complete human virome, including viruses that cause no overt symptoms of disease, but may have unrecognized immunological or developmental consequences. We have used NGS to identify RNA viruses in the blood of 195 patients with UAFI and compared them with those found in 328 apparently healthy (i.e., no overt signs of illness) control individuals, all from communities in southeastern Nigeria. Among UAFI patients, we identified the presence of nucleic acids from several well-characterized pathogenic viruses, such as HIV-1, hepatitis, and Lassa virus. In our cohort of healthy individuals, however, we detected the nucleic acids of two novel rhabdoviruses. These viruses, which we call Ekpoma virus-1 (EKV-1) and Ekpoma virus-2 (EKV-2), are highly divergent, with little identity to each other or other known viruses. The most closely related rhabdoviruses are members of the genus Tibrovirus and Bas-Congo virus (BASV), which was recently identified in an individual with symptoms resembling hemorrhagic fever. Furthermore, by conducting a serosurvey of our study cohort, we find evidence for remarkably high exposure rates to the identified rhabdoviruses. The recent discoveries of novel rhabdoviruses by multiple research groups suggest that human infection with rhabdoviruses might be common. While the prevalence and clinical significance of these viruses are currently unknown, these viruses could have previously unrecognized impacts on human health; further research to understand the immunological and developmental impact of these viruses should be explored. More generally, the identification of similar novel viruses in individuals with and without overt symptoms of disease highlights the need for a broader understanding of the human virome as efforts for viral detection and discovery advance.

Conflict of interest statement

The authors have declared that no competing interests exist.


Fig 1
Fig 1. Viruses identified in UAFI samples and afebrile controls.
(A) Overview of reads we identified in Illumina HiSeq reads from 94 singleton UAFI samples and 34 apparently healthy singleton control samples. For all samples, we removed human reads and common bacterial contaminants, and subjected the remaining reads to BLASTn and BLASTx queries of GenBank and assigned to taxonomic kingdoms using MEGAN 4. (B) Viruses identified in singleton (RNA-seq libraries constructed from a single individual) and pooled libraries (RNA-seq libraries constructed from several individuals). In the case of pooled libraries, the percentage refers to the number of libraries, not individual samples.
Fig 2
Fig 2. Sequencing results and schematic representation of the EKV-1 and -2 genome organization.
(A) Overview of the data generated for each novel rhabdovirus. (B) A schematic showing the assembled genomes, consisting of the following genes: nucleoprotein (N), phosphoprotein (P), matrix (M), U1/U2/U3 (uncharacterized accessory proteins), glycoprotein (G), and polymerase (L). We indicate in orange (EKV-1) and blue (EKV-2) segments of the viral genomes that could not be assembled from Illumina reads and instead Sanger sequenced. (C) Coverage plots of the final viral genomes.
Fig 3
Fig 3. Phylogenetic analysis of rhabdovirus polymerase proteins.
We created a Bayesian phylogenetic tree using full-length polymerase (L) proteins obtained from GenBank. (A) Tree based on alignments of the tibroviruses and ephemeroviruses. Posterior probabilities are shown at relevant nodes (scale bar = nucleotide substitutions/site) and the tree was midpoint rooted. (B) ELISA detection of EKV-1 and EKV-2 IgG in cohorts from Nigeria and the United States (US normals). Each circle correspond to the raw OD450 value of an individual sample. The mean +/- standard deviation (SD) is shown. Black dotted line = cut-off values for positive samples based on the mean of US normals plus 5x or 3x SD. P-values were calculated using a two-tailed Mann-Whitney test.
Fig 4
Fig 4. Examples of rhabdoviruses reported in Africa.
A map depicting examples of rhabdoviruses isolated in sub-Saharan Africa. This map does not depict the current distribution of rhabdoviruses in Sub-Saharan Africa, nor is it meant as a comprehensive listing of all rhabdoviruses isolated in Africa; rather its purpose is to illustrate that many rhabdoviruses have been discovered throughout Africa over the past half-century. Country refers to the sample’s country of origin. Abbreviations: CAR, Central African Republic; DRC, Democratic Republic of Congo.

Similar articles

See all similar articles

Cited by 23 articles

See all "Cited by" articles


    1. Koboldt DC, Steinberg KM, Larson DE, Wilson RK, Mardis ER. The next-generation sequencing revolution and its impact on genomics. Cell. 2013;155: 27–38. 10.1016/j.cell.2013.09.006 - DOI - PMC - PubMed
    1. Chiu CY. Viral pathogen discovery. Curr Opin Microbiol. 2013;16: 468–478. 10.1016/j.mib.2013.05.001 - DOI - PMC - PubMed
    1. Wolfe ND, Dunavan CP, Diamond J. Origins of major human infectious diseases. Nature. 2007;447: 279–283. - PubMed
    1. McMullan LK, Folk SM, Kelly AJ, MacNeil A, Goldsmith CS, Metcalfe MG, et al. A new phlebovirus associated with severe febrile illness in Missouri. N Engl J Med. 2012;367: 834–841. 10.1056/NEJMoa1203378 - DOI - PubMed
    1. Briese T, Paweska JT, McMullan LK, Hutchison SK, Street C, Palacios G, et al. Genetic detection and characterization of Lujo virus, a new hemorrhagic fever-associated arenavirus from southern Africa. PLoS Pathog. 2009;5 e1000455 10.1371/journal.ppat.1000455 - DOI - PMC - PubMed

Publication types

MeSH terms