Design and heterologous expression of dengue virus envelope protein (E) peptides and their use for serological diagnosis

• Published in: Journal of virological methods 2012-12, Vol.186 (1-2), p.55-61
• Main Authors: Honda, Eduardo R., Zanchi, Fernando, Rios, Katiuscia, Lira, Ednaldo, DeusileneVieira, da Silva, Luiz H.P., De Paula, Sergio O.
• Format: Article
• Language: English
• Subjects: antibodies; Antibodies, Viral - blood; antigens; Bioinformatics; Biological and medical sciences; Dengue; Dengue - diagnosis; Dengue virus; Dengue Virus - genetics; Dengue Virus - immunology; Diagnosis; enzyme-linked immunosorbent assay; Flaviviridae; Fundamental and applied biological sciences. Psychology; heterologous gene expression; Humans; immunoglobulin G; Immunoglobulin G - blood; immunoglobulin M; Immunoglobulin M - blood; Indirect ELISA; Microbiology; Peptides; Peptides - genetics; Peptides - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Sensitivity and Specificity; serodiagnosis; Serologic Tests - methods; signs and symptoms (animals and humans); subtropics; Techniques used in virology; Viral Envelope Proteins - genetics; Viral Envelope Proteins - metabolism; Virology; viruses
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2012.08.006

► A definitive diagnosis of dengue infection can be made only in the laboratory and depends on isolating the virus or detecting specific antibodies. ► Currently, the greatest difficulty in conducting immunoenzymatic assays is obtaining suitable antigens. ► We mapped two peptides of the E protein from DENV using bioinformatics tools, expressed these peptides and tested the antigenic potential. ► The results obtained here led to the conclusion that a combination of the E1 and E2 peptides can be used in dengue diagnosis. Viruses belonging to the Flaviviridae family are found and distributed in most of the tropical and sub-tropical regions of the world. The genus has more than 56 members, most of which cause clinical symptoms in humans. The clinical diagnosis of dengue requires laboratory confirmation because of the similarity of symptoms with a series of other acute fevers and the primary use antibodies or antigens for detection. In this work, peptides E1 and E2 of the envelope protein (E) of the dengue virus were mapped using bioinformatics methods. These peptides were then expressed in a prokaryotic system and purified. An indirect ELISA for antibodies IgG and IgM from laboratory samples previously characterised was then used with the peptides to detect anti-dengue antibodies. For IgG using the peptide E1, the sensitivity of the indirect ELISA was 88.3% and the specificity was 56%; using the peptide E2, the sensitivity was 90% and the specificity was 59%; and using a combination of both peptides, the sensitivity was 93.3% and the specificity was 78%. For IgM using the peptide E1, the sensitivity was 88% and the specificity was 66%; using the peptide E2, the sensitivity was 88% and the specificity was 69%; and when used in combination, the peptides E1/E2 demonstrated a sensitivity of 90% and a specificity of 86%. These results indicate that the use of the E1 and E2 peptides of the E protein are an alternative for serological diagnosis of dengue fever.