Nucleoprotein-based indirect enzyme-linked immunosorbent assay (indirect ELISA) for detecting antibodies specific to Ebola virus and Marbug virus
Full-length nucleoproteins from Ebola and Marburg viruses were expressed as His-tagged recombinant proteins in Escherichia coli and nucleoprotein-based enzyme-linked immunosorbent assays (ELISAs) were established for the detection of antibodies specific to Ebola and Marburg viruses. The ELISAs were evaluated by testing antisera collected from rabbit immunized with Ebola and Marburg virus nucleoproteins. Although little cross-reactivity of antibodies was observed in anti-Ebola virus nucleoprotein rabbit antisera, the highest reactions to immunoglobulin G (IgG) were uniformly detected against the nucleoprotein antigens of homologous viruses. We further evaluated the ELISA’s ability to detect antibodies to Ebola and Marburg viruses using human sera samples collected from individuals passing through the Guangdong port of entry. With a threshold set at the mean plus three standard deviations of average optical densities of sera tested, the ELISA systems using these two recombinant nucleoproteins have good sensitivity and specificity. These results demonstrate the usefulness of ELISA for diagnostics as well as ecological and serosurvey studies of Ebola and Marburg virus infection.
KeywordsEbola virus Marburg virus recombinant nucleoproteins indirect ELISA
Unable to display preview. Download preview PDF.
- Barrette R W, Metwally S A, Rowland J M, Xu L, Zaki S R, Nichol S T, Rollin P E, Towner J S, Shieh W J, Batten B, Sealy T K, Carrillo C, Moran K E, Bracht A J, Mayr G A, Sirios-Cruz M, Catbagan D P, Lautner E A, Ksiazek T G, White W R, McIntosh M T. 2009. Discovery of swine as a host for the Reston ebolavirus. Science, 325: 204–206.PubMedCrossRefGoogle Scholar
- Ikegami T, Niikura M, Saijo M, Miranda M E, Calaor A B, Hernandez M, Acosta L P, Manalo D L, Kurane I, Yoshikawa Y, Morikawa S. 2003. Antigen capture enzyme-linked immunosorbent assay for specific detection of Reston Ebola virus nucleoprotein. Clin Diagn Lab Immunol, 10:552–557.PubMedCentralPubMedGoogle Scholar
- Ksiazek T G, Rollin P E, Williams A J, Bressler D S, Martin M L, Swanepoel R, Burt F J, Leman P A, Khan A S, Rowe A K, Mukunu R, Sanchez A, Peters C J. 1999b. Clinical virology of Ebola hemorrhagic fever (EHF): virus, virus antigens, and IgG and IgM antibody findings among EHF patients in Kikwit, Democratic Republic of the Congo, 1995. J Infect Dis, 179:S177–S189.PubMedCrossRefGoogle Scholar
- Kuhn J H, Bao Y M, Bavari S, Becker S, Bradfute S, Brister J R, Bukreyev A A, Cai Y, Chandran K, Davey R A, Dolnik O, Dye J M, Enterlein S, Gonzalez J P, Formenty P, Freiberg A N, Hensley L E, Honko A N, Ignatyev G M, Jahrling P B, Johnson K M, Klenk H D, Kobinger G, Lackemeyer M G, Leroy E M, Lever M S, Lofts L L, Mühlberger E, Netesov S V, Olinger G G, Palacios G, Patterson J L, Paweska J T, Pitt L, Radoshitzky S R, Ryabchikova E I, Saphire E O, Shestopalov A M, Smither S J, Sullivan N J, Swanepoel R, Takada A, Towner J S, van der Groen G, Volchkov V E, Wahl-Jensen V, Warren T K, Warfield K L, Weidmann M, Nichol S T. 2013. Virus nomenclature below the species level: a standardized nomenclature for laboratory animal-adapted strains and variants of viruses assigned to the family Filoviridae. Arch Virol, 158:1425–1432.PubMedCentralPubMedCrossRefGoogle Scholar
- Negredo A, Palacios G, Vazquez-Moron S, Gonzalez F, Dopazo H, Molero F, Juste J, Quetglas J, Savji N, de la Cruz M M, Herrera JE, Pizarro M, Hutchison S K, Echevarria J E, Lipkin W I, Tenorio A. 2011. Discovery of an ebolavirus-like filovirus in europe. PLoS Pathog, 7: e1002304.PubMedCentralPubMedCrossRefGoogle Scholar
- Sanchez A, Geisbert T W, Feldmann H. 2007. In: Filoviridae: Marburg and Ebola viruses. Knipe D M, Howley P M (eds), Fields Virology, 5th ed, Philadelphia: Lippincott Williams and Wilkins, pp 1409–1448.Google Scholar
- Towner J S, Amman B R, Sealy T K, Carroll S A, Comer J A, Kemp A, Swanepoel R, Paddock C D, Balinandi S, Khristova M L, Formenty P B, Albarino C G, Miller D M, Reed Z D, Kayiwa J T, Mills J N, Cannon D L, Greer P W, Byaruhanga E, Farnon E C, Atimnedi P, Okware S, Katongole-Mbidde E, Downing R, Tappero J W, Zaki S R, Ksiazek T G, Nicho S T, Rollin P E. 2009. Isolation of genetically diverse Marburg viruses from Egyptian fruit bats. PLoS Pathog, 5:e1000536.PubMedCentralPubMedCrossRefGoogle Scholar
- WHO: EBOLA RESPONSE ROADMAP UPDATE 26 September 2014. http://apps.who.int/iris/bitstream/10665/135029/1/roadmapupdate26sept14_eng.pdf.