title: Development of an indirect ELISA method for the parallel measurement of IgG and IgM antibodies against Crimean-Congo haemorrhagic fever (CCHF) virus using recombinant nucleoprotein as antigen.
abstract: Recombinant nucleoprotein from Crimean-Congo Haemorrhagic Fever (CCHF) virus was successfully derived from a baculovirus expression system and purified for use in a novel enzyme-linked immunosorbent assay (ELISA) diagnostic test. Comparable tests were used for detection of IgG and IgM antibodies, thus allowing efficient detection of both antibodies in parallel. The major benefits of the assay also included removing any requirement for polyclonal sera, thus eliminating variation in preparations and allowing standardisation between laboratories. The assay was successfully tested using a panel of positive sera supplied from samples identified as being positive in Turkey, Tajikistan and Kosovo and shown to be sensitive and specific. It is envisaged that this simple diagnostic ELISA for CCHF virus infection which removes the reliance on polyclonal antibody preparations, will be accessible to a wider range of laboratories enabling them to carry out routine diagnosis. This will improve the efficiency of diagnosis and subsequent management of infected patients.
authors.affiliation: Health Protection Agency, Porton Down, Salisbury, Wiltshire SP4 0JG, UK. stuart.dowall@hpa.org.uk
authors.forename: S D
authors.initials: SD
authors.lastname: Dowall
authors.forename: K S
authors.initials: KS
authors.lastname: Richards
authors.forename: V A
authors.initials: VA
authors.lastname: Graham
authors.forename: J
authors.initials: J
authors.lastname: Chamberlain
authors.forename: R
authors.initials: R
authors.lastname: Hewson
mesh_terms: D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics; D016367:Baculoviridae / Q000235:genetics; D019411:Clinical Laboratory Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005822:Genetic Vectors; D003225:Hemorrhagic Fever Virus, Crimean-Congo / Q000276:immunology*; D006479:Hemorrhagic Fever, Crimean / Q000175:diagnosis*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D009698:Nucleoproteins / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D013620:Tajikistan; D014421:Turkey; D015022:Yugoslavia
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D009698:Nucleoproteins; D011994:Recombinant Proteins