title: Expression, purification, and evaluation of diagnostic potential and immunogenicity of a recombinant NS3 protein from all serotypes of dengue virus.
abstract: Dengue is one of the major public health concerns in the world. Since all the four serotypes are actively circulating in Mexico, there is a need to develop an efficient diagnosis system to improve case management of the patients. There exist few studies evaluating the use of the NS3 protein as a protective antigen against dengue virus (DENV). In this paper we show the expression of a recombinant NS3 protein from all serotypes of dengue virus (GST-DVNS3-1-4) and report a reliable "in-house detection system" for the diagnosis of dengue infection which was field-tested in a small village (Tezonapa) in the state of Veracruz, Mexico. The fusion proteins were immunogenic, inducing antibodies to be able to recognize to antigens up to a 1 : 3200 dilution. The purified proteins were used to develop an in-house detection system (ELISA) and were further tested with a panel of 239 serum samples. The in-house results were in excellent agreement with the commercial kits with κ = 0.934 ± 0.064 (95%  CI = 0.808-1.061), and κ = 0.872 ± 0.048 (95%  CI = 0.779-0.965) for IgM and IgG, respectively. The agreement between the NS1 antigen detection versus the rNS3 ELISA, κ = 0.837 ± 0.066 (95%  CI = 0.708-0.966), was very good. Thus, these results demonstrate that recombinant NS3 proteins have potential in early diagnosis of dengue infections.
authors.affiliation: LADISER Inmunología y Biología Molecular, Facultad de Ciencias Químicas, Universidad Veracruzana, 94340 Orizaba, VER, Mexico ; Centro de Investigaciones Biomédicas, Universidad Veracruzana, 91190 Xalapa, VER, Mexico.
authors.forename: Laura Mónica
authors.initials: LM
authors.lastname: Alvarez-Rodríguez
authors.forename: Angel
authors.initials: A
authors.lastname: Ramos-Ligonio
authors.forename: José Luis
authors.initials: JL
authors.lastname: Rosales-Encina
authors.forename: María Teresa
authors.initials: MT
authors.lastname: Martínez-Cázares
authors.forename: Aurora
authors.initials: A
authors.lastname: Parissi-Crivelli
authors.forename: Aracely
authors.initials: A
authors.lastname: López-Monteon