Using the remaining uL of BASV-positive serum, further attempts were made to culture the virus in 5 cell lines and in suckling mice. Pups were observed daily for 14 days for lethality or signs of clinical illness.

The second diluted sample was inoculated into Since no CPE was observed in any of the cultures, cells were subsequently fixed for transmission electron microscopy to see if viral particles could be visualized [45]. The culture medium was replaced 15 hours post-transfection and cells were stimulated with 6.

The inoculum was removed after 4 hours and replaced by fresh culture medium. At 24 hours post-infection, infectious supernatants were harvested, filtered through 0. Each phylogenetic tree is rooted by using the corresponding protein from human parainfluenza virus type 1 HPIV-1 , a paramyxovirus, as an outgroup.

Rotavirus is readily detected in extracted RNA from a stool sample taken from an ongoing study of viral diarrhea in the laboratory lane 1 , but not in two separate aliquots of extracted nucleic acid from the BASV serum sample lanes 2 and 3. Viral reads in the deep sequencing data corresponding to the BASV-positive serum sample.

The authors thank the national and international teams involved in the control of suspected hemorrhagic fever cases that occurred in in Democratic Republic of the Congo DRC.

Obtained consents from patients and their families: PM. Introduction Viral hemorrhagic fever VHF encompasses a group of diseases characterized by fever, malaise, bleeding abnormalities, and circulatory shock [1] , [2] , [3]. Download: PPT. Figure 1. Map of Africa showing countries that are affected by viral hemorrhagic fever VHF outbreaks.

Table 1. Demographics of and clinical symptoms developed in the three patients suspected to be infected by Bas-Congo virus BASV. Patient 2. Patient 3. Initial Sample Collection and Diagnostic Testing A cluster of three human cases of typical acute hemorrhagic fever occurred between May 25 and June 13, in Mangala village, located in a remote tropical forest region in Central Africa.

Discovery and Genome Assembly of the BASV Rhabdovirus To identify a potential causative pathogen in the third surviving patient with unknown hemorrhagic fever, RNA extracts from the serum sample were analyzed using unbiased deep sequencing Fig.

Figure 2. Deep sequencing and whole-genome de novo assembly of BASV. Phylogenetic Analysis of BASV and Comparison with other Rhabdoviruses Phylogenetic trees reveal that BASV belongs to the dimarhabdoviridae supergroup and is distantly related to members of the Tibrogargan group and the Ephemerovirus genus, although it clusters separately from other rhabdoviruses in an independent deeply rooted branch Figs.

Figure 3. Figure 4. Figure 5. Schematic representation of the genome organization of BASV and its protein similarity plot compared to representative rhabdoviruses.

Figure 6. Discussion Among more than species of rhabdoviruses identified to date, fewer than 10 have been isolated from humans [24]. Figure 8. Methods Ethics Statement Written informed consent for publication of their case reports was obtained from the sole survivor of the hemorrhagic fever outbreak and the parents of the two deceased children.

Diagnostic Samples No diagnostic samples were available from Patient 1 or Patient 2. Supporting Information. Figure S1. Figure S2. Table S1. Table S2. Acknowledgments The authors thank the national and international teams involved in the control of suspected hemorrhagic fever cases that occurred in in Democratic Republic of the Congo DRC. References 1. Bray M Pathogenesis of viral hemorrhagic fever. Curr Opin Immunol — View Article Google Scholar 2. Hum Vaccin 7: — View Article Google Scholar 3.

Nat Med S— View Article Google Scholar 4. JAMA — View Article Google Scholar 5. View Article Google Scholar 6. Ergonul O Crimean-Congo haemorrhagic fever.

Lancet Infect Dis 6: — View Article Google Scholar 7. Clin Microbiol Infect — View Article Google Scholar 8. Monath TP Dengue and yellow fever—challenges for the development and use of vaccines. N Engl J Med — View Article Google Scholar 9. Infect Genet Evol 9: — View Article Google Scholar Fu ZF Genetic comparison of the rhabdoviruses from animals and plants.

Curr Top Microbiol Immunol 1— Indian J Med Res — Lancet — Virus Res — Tang P, Chiu C Metagenomics for the discovery of novel human viruses. Future Microbiol 5: — PLoS Pathog 5: e Science — PLoS One 5: e J Virol — Virology 13— Springfeld C, Darai G, Cattaneo R Characterization of the Tupaia rhabdovirus genome reveals a long open reading frame overlapping with P and a novel gene encoding a small hydrophobic protein. J Gen Virol — Virus Res 17— Fu ZF The world of rhabdoviruses.

Berlin; New York: Springer. Am J Trop Med Hyg — Vital signs: incidence and trends of infection with pathogens transmitted commonly through food—foodborne diseases active surveillance network, 10 U. Deanna Lee Search articles by 'Deanna Lee'. Lee D ,. Elizabeth Slikas Search articles by 'Elizabeth Slikas'. Slikas E ,. Steffen I ,. Muyembe JJ ,. Sittler T ,. Narayanan Veeraraghavan Search articles by 'Narayanan Veeraraghavan'. Veeraraghavan N ,. Ruby JG ,.

Chunlin Wang Search articles by 'Chunlin Wang'. Wang C ,. Maria Makuwa Search articles by 'Maria Makuwa'. Makuwa M ,. Prime Mulembakani Search articles by 'Prime Mulembakani'.

Mulembakani P ,. Tesh RB ,. Jonna Mazet Search articles by 'Jonna Mazet'. Mazet J ,. Rimoin AW ,. Travis Taylor Search articles by 'Travis Taylor'. Taylor T ,. Schneider BS ,. Simmons G ,. Delwart E ,. Wolfe ND ,. Chiu CY ,. Leroy EM. Show less. Affiliations 1 author 1. This article has been corrected. See PLoS Pathog. Share this article Share with email Share with twitter Share with linkedin Share with facebook. Free full text. PLoS Pathog. Published online Sep PMID: Gilda Grard , 1 , 2 Joseph N.

Tesh , 11 Jonna Mazet , 12 Anne W. Rimoin , 13 Travis Taylor , 3 Bradley S. Wolfe , 3 Charles Y. Joseph N. Graham Ruby. Robert B. Anne W. Bradley S. Nathan D. Charles Y. Eric M. David Wang, Editor. Author information Article notes Copyright and License information Disclaimer. The authors have filed a patent application related to BASV. This does not alter the authors' adherence to all PLOS Pathogens policies on sharing data and materials.

Received May 23; Accepted Aug 8. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

This article has been cited by other articles in PMC. Go to:. Open in a separate window. Figure 1. Map of Africa showing countries that are affected by viral hemorrhagic fever VHF outbreaks. Case Reports from an Acute Hemorrhagic Fever Outbreak Patient 1 The first case was a year-old boy who presented to the health center in Mangala village Boma Bungu Health Zone on May 25, with malaise, epistaxis nose bleeding , conjunctival injection, gingival bleeding, hematemesis vomiting with blood , and watery diarrhea with blood Table 1.

Patient 2 The second case was a year-old girl. Patient 3 The third case was a male nurse aged 32 years working in the health center visited by Patients 1 and 2.

Initial Sample Collection and Diagnostic Testing A cluster of three human cases of typical acute hemorrhagic fever occurred between May 25 and June 13, in Mangala village, located in a remote tropical forest region in Central Africa.

Discovery and Genome Assembly of the BASV Rhabdovirus To identify a potential causative pathogen in the third surviving patient with unknown hemorrhagic fever, RNA extracts from the serum sample were analyzed using unbiased deep sequencing Fig. Figure 2. Deep sequencing and whole-genome de novo assembly of BASV. Phylogenetic Analysis of BASV and Comparison with other Rhabdoviruses Phylogenetic trees reveal that BASV belongs to the dimarhabdoviridae supergroup and is distantly related to members of the Tibrogargan group and the Ephemerovirus genus, although it clusters separately from other rhabdoviruses in an independent deeply rooted branch Figs.

Figure 3. Figure 4. Figure 5. Schematic representation of the genome organization of BASV and its protein similarity plot compared to representative rhabdoviruses.

Figure 6. Figure 7. Figure 8. Ethics Statement Written informed consent for publication of their case reports was obtained from the sole survivor of the hemorrhagic fever outbreak and the parents of the two deceased children. Diagnostic Samples No diagnostic samples were available from Patient 1 or Patient 2. TIF Click here for additional data file. DOCX Click here for additional data file. Bray M Pathogenesis of viral hemorrhagic fever. Curr Opin Immunol 17 : — Hum Vaccin 7 : — Nat Med 10 : S— JAMA : — Ergonul O Crimean-Congo haemorrhagic fever.

Lancet Infect Dis 6 : — Clin Microbiol Infect 17 : — Monath TP Dengue and yellow fever—challenges for the development and use of vaccines. N Engl J Med : — Infect Genet Evol 9 : — Fu ZF Genetic comparison of the rhabdoviruses from animals and plants. Curr Top Microbiol Immunol : 1— Indian J Med Res : — Lancet : — Virus Res 85 : — Tang P, Chiu C Metagenomics for the discovery of novel human viruses.

Future Microbiol 5 : — PLoS Pathog 5 : e Science : — PLoS One 5 : e J Virol 78 : — Virus Res : — Virology : 13— Springfeld C, Darai G, Cattaneo R Characterization of the Tupaia rhabdovirus genome reveals a long open reading frame overlapping with P and a novel gene encoding a small hydrophobic protein.

J Virol 79 : — J Gen Virol 92 : — Virus Res : 17— Fu ZF The world of rhabdoviruses. Berlin; New York: Springer. Am J Trop Med Hyg 68 : — Vital signs: incidence and trends of infection with pathogens transmitted commonly through food—foodborne diseases active surveillance network, 10 U. Pediatr Infect Dis J 21 : — Emerg Infect Dis 11 : — Curr Top Microbiol Immunol : 81— Imported Lassa fever—New Jersey, J Clin Microbiol 45 : — Stanifer ML, Cureton DK, Whelan SP A recombinant vesicular stomatitis virus bearing a lethal mutation in the glycoprotein gene uncovers a second site suppressor that restores fusion.

J Virol 85 : — Samples from the lucky nurse have been tested and it turned out a completely new virus had infected him, Fair and other researchers report in the Public Library of Science journal PLoS pathogens. The genetic sequences went to Dr. Yet about 20 percent of the time we have hemorrhagic fever outbreaks that are completely negative, which means unknown causes and they are not Ebola.

The sequencing puts this new virus on its own branch of the bad virus family tree -- somewhat related to Ebola and the virus that causes Lassa fever, another horrific killer, and most closely related to the rhabdoviruses. This family usually only infects animals with one notable exception -- rabies. But rabies is not known to cause hemorrhaging. A nurse who took care of the first infected nurse had antibodies to the new virus. Tests of other villagers have found no more evidence of the virus, however, which is good news.

Africa is loaded with nasty viruses. Lassa fever virus comes from a family known as arenaviruses and causes , cases of hemorrhagic fever a year. Crimean-Congo hemorrhagic fever and Rift Valley Fever viruses are in another family called bunyaviruses; Ebola and Marburg viruses are filoviruses that kill anywhere between 30 percent and 90 percent of victims.

This adds a new one to the list.