Enzyme-linked Immunosorbent Assays for Detection of Antibodies to Ebola and Marburg Viruses Using Recombinant Nucleoproteins

Overview

Journal J Clin Microbiol

Specialty Microbiology

Date 2001 Jan 4

PMID 11136739

Citations 37

Authors

M Saijo

M Niikura

S Morikawa

T G Ksiazek

R F Meyer

C J Peters

I Kurane

Affiliations

Soon will be listed here.

Abstract

The full-length nucleoprotein (NP) of Ebola virus (EBO) was expressed as a His-tagged recombinant protein (His-EBO-NP) by a baculovirus system. Carboxy-terminal halves of NPs of EBO and Marburg virus (MBG) were expressed as glutathione S-transferase-tagged recombinant proteins in an Escherichia coli system. The antigenic regions on the NPs of EBO and MBG were determined by both Western blotting and enzyme-linked immunosorbent assay (ELISA) to be located on the C-terminal halves. The C-terminal 110 and 102 amino acids of the NPs of EBO and MBG, respectively, possess strong antigenicity. The full-length NP of EBO was strongly expressed in insect cells upon infection with the recombinant baculovirus, while expression of the full-length NP of MBG was weak. We developed an immunoglobulin G (IgG) ELISA using His-EBO-NP and the C-terminal halves of the NPs of EBO and MBG as antigens. We evaluated the IgG ELISA for the ability to detect IgG antibodies to EBO and MBG, using human sera collected from EBO and MBG patients. The IgG ELISA with the recombinant NPs showed high sensitivity and specificity in detecting EBO and MBG antibodies. The results indicate that ELISA systems prepared with the recombinant NPs of EBO and MBG are valuable tools for the diagnosis of EBO and MBG infections and for seroepidemiological field studies.

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References

van der Groen G, Kurata T, METS C . Modifications to indirect immunofluorescence tests on Lassa, Marburg, and Ebola material. Lancet. 1983; 1(8325):654. DOI: 10.1016/s0140-6736(83)91831-7. View

Siegert R, SHU H, Slenczka W, Peters D, Muller G . [On the etiology of an unknown human infection originating from monkeys]. Dtsch Med Wochenschr. 1967; 92(51):2341-3. DOI: 10.1055/s-0028-1106144. View

Becker S, Huppertz S, Klenk H, Feldmann H . The nucleoprotein of Marburg virus is phosphorylated. J Gen Virol. 1994; 75 ( Pt 4):809-18. DOI: 10.1099/0022-1317-75-4-809. View

Saijo M, Suzutani T, Itoh K, Hirano Y, Murono K, Nagamine M . Nucleotide sequence of thymidine kinase gene of sequential acyclovir-resistant herpes simplex virus type 1 isolates recovered from a child with Wiskott-Aldrich syndrome: evidence for reactivation of acyclovir-resistant herpes simplex virus. J Med Virol. 1999; 58(4):387-93. View

Bertherat E, Talarmin A, Zeller H . [Democratic Republic of the Congo: between civil war and the Marburg virus. International Committee of Technical and Scientific Coordination of the Durba Epidemic]. Med Trop (Mars). 1999; 59(2):201-4. View

Smith C, Simpson D, BOWEN E, ZLOTNIK I . Fatal human disease from vervet monkeys. Lancet. 1967; 2(7526):1119-21. DOI: 10.1016/s0140-6736(67)90621-6. View

Feldmann H, Slenczka W, Klenk H . Emerging and reemerging of filoviruses. Arch Virol Suppl. 1996; 11:77-100. DOI: 10.1007/978-3-7091-7482-1_9. View

Muyembe-Tamfum J, Kipasa M, Kiyungu C, Colebunders R . Ebola outbreak in Kikwit, Democratic Republic of the Congo: discovery and control measures. J Infect Dis. 1999; 179 Suppl 1:S259-62. DOI: 10.1086/514302. View

Prehaud C, Hellebrand E, Coudrier D, Volchkov V, Volchkova V, Feldmann H . Recombinant Ebola virus nucleoprotein and glycoprotein (Gabon 94 strain) provide new tools for the detection of human infections. J Gen Virol. 1998; 79 ( Pt 11):2565-72. DOI: 10.1099/0022-1317-79-11-2565. View

10.

Jahrling P, Geisbert T, Dalgard D, Johnson E, Ksiazek T, Hall W . Preliminary report: isolation of Ebola virus from monkeys imported to USA. Lancet. 1990; 335(8688):502-5. DOI: 10.1016/0140-6736(90)90737-p. View

11.

Johnson K, ELLIOTT L, Heymann D . Preparation of polyvalent viral immunofluorescent intracellular antigens and use in human serosurveys. J Clin Microbiol. 1981; 14(5):527-9. PMC: 273981. DOI: 10.1128/jcm.14.5.527-529.1981. View

12.

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

13.

Muyembe T, Kipasa M . Ebola haemorrhagic fever in Kikwit, Zaire. International Scientific and Technical Committee and WHO Collaborating Centre for Haemorrhagic Fevers. Lancet. 1995; 345(8962):1448. DOI: 10.1016/s0140-6736(95)92640-2. View

14.

Sanchez A, Kiley M, Holloway B, McCormick J, Auperin D . The nucleoprotein gene of Ebola virus: cloning, sequencing, and in vitro expression. Virology. 1989; 170(1):81-91. DOI: 10.1016/0042-6822(89)90354-1. View

15.

Kitts P, Ayres M, Possee R . Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors. Nucleic Acids Res. 1990; 18(19):5667-72. PMC: 332298. DOI: 10.1093/nar/18.19.5667. View

16.

Matsuura Y, Possee R, Overton H, Bishop D . Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. J Gen Virol. 1987; 68 ( Pt 5):1233-50. DOI: 10.1099/0022-1317-68-5-1233. View

17.

Ksiazek T, Rollin P, Williams A, Bressler D, Martin M, Swanepoel R . Clinical virology of Ebola hemorrhagic fever (EHF): virus, virus antigen, and IgG and IgM antibody findings among EHF patients in Kikwit, Democratic Republic of the Congo, 1995. J Infect Dis. 1999; 179 Suppl 1:S177-87. DOI: 10.1086/514321. View

18.

Johnson K, Lange J, WEBB P, Murphy F . Isolation and partial characterisation of a new virus causing acute haemorrhagic fever in Zaire. Lancet. 1977; 1(8011):569-71. DOI: 10.1016/s0140-6736(77)92000-1. View

19.

. Ebola haemorrhagic fever in Sudan, 1976. Report of a WHO/International Study Team. Bull World Health Organ. 1978; 56(2):247-70. PMC: 2395561. View

20.

BOWEN E, Lloyd G, Harris W, Platt G, Baskerville A, Vella E . Viral haemorrhagic fever in southern Sudan and northern Zaire. Preliminary studies on the aetiological agent. Lancet. 1977; 1(8011):571-3. DOI: 10.1016/s0140-6736(77)92001-3. View