Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E-coli enable sensitive and early dengue diagnosis

被引：23

作者：

Allonso, Diego ^{[1
]}

Rosa, Marcela da Silva ^{[1
]}

Coelho, Diego Rodrigues ^{[1
]}

da Costa, Simone Morais ^{[2
]}

Ribeiro Nogueira, Rita Maria ^{[3
]}

Bozza, Fernando Augusto ^{[4
]}

Dos Santos, Flavia Barreto ^{[3
]}

de Barcelos Alves, Ada Maria ^{[2
]}

Mohana-Borges, Ronaldo ^{[1
]}

机构：

[1] Univ Fed Rio de Janeiro, Inst Biofis Carlos Chagas Filho, Lab Genom Estruct, BR-21941590 Rio De Janeiro, Brazil

[2] Fundacao Oswaldo Cruz, Inst Oswaldo Cruz, Lab Biotecnol & Fisiol Infeccoes Virais, Rio De Janeiro, Brazil

[3] Fundacao Oswaldo Cruz, Inst Oswaldo Cruz, Lab Flavivirus, Rio De Janeiro, Brazil

[4] Fundacao Oswaldo Cruz, Inst Pesquisas Clin Evandro Chagas, Unidade Terapia Intens, Rio De Janeiro, Brazil

来源：

JOURNAL OF VIROLOGICAL METHODS | 2011年 / 175卷 / 01期

关键词：

Dengue virus; NS1; protein; Dengue diagnosis; E; coli; NONSTRUCTURAL GLYCOPROTEIN NS1; SECONDARY STRUCTURE ANALYSES; ANTIGEN CAPTURE IMMUNOASSAY; LINKED-IMMUNOSORBENT-ASSAY; MONOCLONAL-ANTIBODIES; SIGNAL SEQUENCE; DNA VACCINE; TYPE-1; INFECTIONS; CELLS;

D O I：

10.1016/j.jviromet.2011.04.029

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The non-structural 1 (NS1) protein plays an important role in dengue diagnosis because it has been detected as a soluble serum antigen in both primary and secondary infections. The NS1 protein was expressed in Escherichia coli cells, and the efficiency of four different refolding protocols was tested. All of the protocols generated dimeric NS1 in a conformation similar to that of the protein expressed by eukaryotic cells. A polyclonal antibody produced from the properly folded E. coli recombinant NS1 (rNS1) protein proved to be a useful tool for the diagnosis of Dengue virus because it detected 100% of the Dengue virus 2 (DENV2) in infected patients' sera and 60% of the DENV IgM-positive sera not detected by commercial NS1-based diagnostic kits. These data suggest a high-efficiency method for correctly folding rNS1 that maintains its structural and immunogenic properties. In addition, a detection method using the polyclonal antibody against correctly folded rNS1 seemed to be more sensitive and efficient for NS1 detection in serum, highlighting its usefulness for developing a high-sensitivity diagnostic kit. (C) 2011 Elsevier B.V. All rights reserved.

引用

页码：109 / 116

页数：8

共 34 条

[1] Enzyme-linked immunosorbent assay specific to dengue virus type 1 nonstructural protein NS1 reveals circulation of the antigen in the blood during the experiencing primary acute phase of disease in patients or secondary infections [J].