title: Expression of non-conserved regions of the S genome segments of three hantaviruses: evaluation of the expressed polypeptides for diagnosis of haemorrhagic fever with renal syndrome.
abstract: Haemorrhagic fever with renal syndrome (HFRS) is a serious and often fatal disease caused by viruses in the Hantavirus genus of the family Bunyaviridae. We expressed the entire coding region of the small (S) genome segments of three serologically distinct hantaviruses as soluble proteins in Escherichia coli and evaluated the expressed nucleocapsid proteins (NPs) as antigens for diagnosis of HFRS. We also prepared novel diagnostic antigens by expressing truncated genes from which we deleted amino acid coding regions that were highly conserved among the three viruses. These antigens were analysed for their potential to detect and differentiate between antisera to various hantaviruses by ELISA. ELISA results obtained with HFRS patient sera or with sera from naturally or experimentally infected animals indicate that homologous antigens and antisera reacted to high titre. The truncated NPs were more specific than the complete NPs in distinguishing between possible aetiological agents of HFRS. Our findings demonstrate that prokaryotic expression of portions of the NPs of specific hantaviruses can be used to generate, readily and efficiently, large quantities of antigen that is both sensitive and specific in diagnostic assays for HFRS.
authors.affiliation: Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 21702-5011.
authors.forename: M
authors.initials: M
authors.lastname: Wang
authors.forename: C
authors.initials: C
authors.lastname: Rossi
authors.forename: C S
authors.initials: CS
authors.lastname: Schmaljohn
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000191:economics / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D005069:Evaluation Studies as Topic; D014644:Genetic Variation; D005822:Genetic Vectors / Q000235:genetics; D006476:Orthohantavirus / Q000145:classification / Q000235:genetics; D006480:Hemorrhagic Fever with Renal Syndrome / Q000175:diagnosis* / Q000453:epidemiology; D006801:Humans; D007674:Kidney Diseases / Q000175:diagnosis* / Q000453:epidemiology; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D016133:Polymerase Chain Reaction; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D011994:Recombinant Proteins; D014758:Viral Core Proteins