title: Expression of a hantavirus N protein and its efficacy as antigen in immune assays.
abstract: Hantavirus cardiopulmonary syndrome (HCPS) has been recognized as an important public heath problem. Five hantaviruses associated with HCPS are currently known in Brazil: Juquitiba, Araraquara, Laguna Negra-like, Castelo dos Sonhos, and Anajatuba viruses. The laboratory diagnosis of HCPS is routinely carried out by the detection of anti-hantavirus IgM and/or IgG antibodies. The present study describes the expression of the N protein of a hantavirus detected in the blood sample of an HCPS patient. The entire S segment of the virus was amplified and found to be 1858 nucleotides long, with an open reading frame of 1287 nucleotides that encodes a protein of 429 amino acids. The nucleotide sequence described here showed a high identity with the N protein gene of Araraquara virus. The entire N protein was expressed using the vector pET200D and the Escherichia coli BL21 strain. The expression of the recombinant protein was confirmed by the detection of a 52-kDa protein by Western blot using a pool of human sera obtained from HCPS patients, and by specific IgG detection in five serum samples of HCPS patients tested by ELISA. These results suggest that the recombinant N protein could be used as an antigen for the serological screening of hantavirus infection.
authors: Centro de Pesquisa em Virologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil. ltmfigue@fmrp.usp.br
authors: L T M
authors: LT
authors: Figueiredo
authors: M L
authors: ML
authors: Moreli
authors: A A
authors: AA
authors: Borges
authors: G G
authors: GG
authors: Figueiredo
authors: R L M
authors: RL
authors: Souza
authors: V H
authors: VH
authors: Aquino
mesh_terms: D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D036022:Capsid Proteins / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D005822:Genetic Vectors; D006476:Orthohantavirus / Q000276:immunology*; D018804:Hantavirus Pulmonary Syndrome / Q000175:diagnosis*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology; D014758:Viral Core Proteins / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D007074:Immunoglobulin G; D019590:Nucleocapsid Proteins; D014758:Viral Core Proteins; C111782:nucleocapsid protein, Hantaan virus