Expression of a hantavirus N protein and its efficacy as antigen in immune assays.
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.
Figueiredo
L T M
LT
Centro de Pesquisa em Virologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil. ltmfigue@fmrp.usp.br
Moreli
M L
ML
Borges
A A
AA
Figueiredo
G G
GG
Souza
R L M
RL
Aquino
V H
VH
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
D000956:Antigens, Viral; D036022:Capsid Proteins; D007074:Immunoglobulin G; D019590:Nucleocapsid Proteins; D014758:Viral Core Proteins; C111782:nucleocapsid protein, Hantaan virus