title: Recombinant VP9-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Banna virus (genus Seadornavirus).
abstract: Banna virus (BAV, genus Seadornavirus, family Reoviridae) is an arbovirus suspected to be responsible for encephalitis in humans. Two genotypes of this virus are distinguishable: A (Chinese isolate, BAV-Ch) and B (Indonesian isolate, BAV-In6969) which exhibit only 41% amino-acid identity in the sequence of their VP9. The VP7 to VP12 of BAV-Ch and VP9 of BAV-In6969 were expressed in bacteria using pGEX-4T-2 vector. VP9 was chosen to establish an ELISA for BAV, based mainly on two observations: (i). VP9 is a major protein in virus-infected cells and is a capsid protein (ii). among all the proteins expressed, VP9 was obtained in high amount and showed the highest immuno-reactivity to anti-BAV ascitic fluid. The VP9s ELISA was evaluated in three populations: French blood donors and two populations (blood donors and patients with a neurological syndrome) from Malaysia, representing the region where the virus was isolated in the past. The specificity of this ELISA was >98%. In mice injected with live BAV, the assay detected IgG-antibody to BAV infection 21 days post-injection, which was confirmed by Western blot using BAV-infected cells. The VP9 ELISA permits to determine the sero-status of a population without special safety precautions and without any requirements to propagate the BAV. This test should be a useful tool for epidemiological survey of BAV.
authors.affiliation: Unité des Virus Emergents (EA 3292, IFR 48): Université de la Méditerranée, Faculté de Médecine de Marseille and EFS Alpes-Méditerranée, Marseille 13005, France.
authors.forename: Fauziah
authors.initials: F
authors.lastname: Mohd Jaafar
authors.forename: Houssam
authors.initials: H
authors.lastname: Attoui
authors.forename: Pierre
authors.initials: P
authors.lastname: Gallian
authors.forename: Ilina
authors.initials: I
authors.lastname: Isahak
authors.forename: Kum Thong
authors.initials: KT
authors.lastname: Wong
authors.forename: Soon Keng
authors.initials: SK
authors.lastname: Cheong
authors.forename: Veera Sekaran
authors.initials: VS
authors.lastname: Nadarajah
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Cantaloube
authors.forename: Philippe
authors.initials: P
authors.lastname: Biagini
authors.forename: Philippe
authors.initials: P
authors.lastname: De Micco
authors.forename: Xavier
authors.initials: X
authors.lastname: De Lamballerie
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D001202:Ascitic Fluid / Q000276:immunology / Q000821:virology; D018162:Coltivirus / Q000276:immunology*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005838:Genotype; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D011994:Recombinant Proteins / Q000276:immunology; D012088:Reoviridae Infections / Q000175:diagnosis* / Q000453:epidemiology; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014764:Viral Proteins