Recombinant VP9-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Banna virus (genus Seadornavirus).
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.
Mohd Jaafar
Fauziah
F
Unité des Virus Emergents (EA 3292, IFR 48): Université de la Méditerranée, Faculté de Médecine de Marseille and EFS Alpes-Méditerranée, Marseille 13005, France.
Attoui
Houssam
H
Gallian
Pierre
P
Isahak
Ilina
I
Wong
Kum Thong
KT
Cheong
Soon Keng
SK
Nadarajah
Veera Sekaran
VS
Cantaloube
Jean-François
JF
Biagini
Philippe
P
De Micco
Philippe
P
De Lamballerie
Xavier
X
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*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014764:Viral Proteins