[An immunoenzyme assay for the detection of monoclonal antibodies against the E and NS1 proteins of the dengue virus].
The ELISA was standardized to detect monoclonal antibodies of dengue virus proteins E and NS1. One indirect ELISA was applied, using C6-36 cells inoculated with the A-15 strain, isolated during the dengue 2 epidemic in 1981 as an antigen source. These cells were fixed in ELISA plates at a 200,000 cell/well concentration. A cell control in similar conditions was used. Specific monoclonal antibodies to both proteins were used to standardize the system. Studies at different incubation periods, to determine the highest expression moment of these proteins in the cell membrane, were carried out. The results show a full response at 72 hours postinoculation for both proteins; a 14.7 ng/mL sensitivity was obtained for the detection of NS1, and of 1.43 ng/mL for E protein. This system allows the monoclonal antibodies primary screening to dengue 2 virus E and NS1 proteins.
Vázquez
S
S
Instituto de Medicina Tropical Pedro Kourí.
Pupo
M
M
Pelegrino
J L
JL
Morier
L
L
Castillo
A
A
Otero
A
A
Guzmán
M G
MG
D000330:Aedes; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000914:Antibodies, Viral / Q000032:analysis*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology*; D002478:Cells, Cultured; D003716:Dengue Virus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D013997:Time Factors; D014759:Viral Envelope Proteins / Q000276:immunology*; D017361:Viral Nonstructural Proteins / Q000276:immunology*; D014764:Viral Proteins / Q000276:immunology*; D014776:Virus Cultivation / Q000379:methods
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; C051821:E-glycoprotein, Dengue virus type 2; C079340:NS1 protein, Dengue virus type 2; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins