Recombinant VP9-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Banna virus (genus Seadornavirus)

• Published in: Journal of virological methods 2004-03, Vol.116 (1), p.55-61
• Main Authors: Jaafar, Fauziah Mohd, Attoui, Houssam, Gallian, Pierre, Isahak, Ilina, Wong, Kum Thong, Cheong, Soon Keng, Nadarajah, Veera Sekaran, Cantaloube, Jean-François, Biagini, Philippe, De Micco, Philippe, De Lamballerie, Xavier
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Viral - blood; Antigens, Viral - genetics; Antigens, Viral - immunology; Ascitic Fluid - immunology; Ascitic Fluid - virology; Banna virus; Banna virus VP9; Biological and medical sciences; Coltivirus - immunology; Cross Reactions; Diagnosis; ELISA; Enzyme-Linked Immunosorbent Assay - methods; Fundamental and applied biological sciences. Psychology; Genotype; Humans; Immunoglobulin G - blood; Mice; Microbiology; Recombinant protein; Recombinant Proteins - immunology; Reoviridae Infections - diagnosis; Reoviridae Infections - epidemiology; Seadornavirus; Sensitivity and Specificity; Techniques used in virology; Viral Proteins - genetics; Viral Proteins - immunology; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2003.10.010

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.