Concanavalin A- Sandwich-ELISA for the detection of antibodies against Newcastle disease virus (NDV) in chicken
KRISHI: Publication and Data Inventory Repository
View Archive InfoField | Value | |
Title |
Concanavalin A- Sandwich-ELISA for the detection of antibodies against Newcastle disease virus (NDV) in chicken
|
|
Creator |
T.R. Kannaki, E. Priyanka and Santosh Haunshi
|
|
Subject |
Chicken
Newcastle Disease Con A |
|
Description |
Not Available
Concanavalin A (Con A), a lectin interacts with carbohydrate moieties of viruses and provide stable and sensitive detection when used as a capture agent. Indirect ELISA methods need purified Newcastle disease virus (NDV) or recombinant antigens for adsorption, whereas use of Con A as capture agent will enable the use of non-purified and non-concentrated virus as antigen replacing costly and time-consuming virus purification step. Con A based sandwich ELISA with nonpurified NDV whole virus antigen with single serum dilution format has been developed in this study. The optimum concentrations of the capture agent, Con A and non-purified antigen preparations were determined by checker-board titration. Briefly, microplates were coated with predetermined optimum concentration of ConA (0.5 mg/ml; 50μg per well) and incubated for 18h at 4C. After washing, allantoic fluid with Newcastle disease virus (NDV) LaSota (HA titre, 210) at a constant predetermined dilution (1:1; 50μl) was coated and incubated for 45 min at 37C, followed by blocking with 2 % bovine serum albumin for 45 min at 37C. The antigen coated plates were used in the detection of antibody titre against NDV in serum samples at single serum dilution of 1: 500. Then, wells were added with goat anti-chicken IgG horseradish peroxidase conjugate and incubated for 1h at 37C, followed by addition of TMB substrate and the plates were read spectrophotometrically at 450 nm. ELISA antibody titres were determined by standard serial dilution of positive sera and endpoints were calculated by a subtraction method. By using positive negative threshold curve (PNT), intercept and slope of the standard curve were calculated. Total of 271 random chicken serum samples were analyzed for antibodies against NDV by Haemagglutination inhibition assay (HI), indirect ELISA and compared with the Con A- S- ELISA developed in this study. The Con A-S-ELISA showed a high coefficient of correlation (r=0.85, n=271, P Not Available |
|
Date |
2019-12-05T05:23:37Z
2019-12-05T05:23:37Z 2019-01-31 |
|
Type |
Research Paper
|
|
Identifier |
Not Available
DOI: 10.18805/ijar.B-3711 http://krishi.icar.gov.in/jspui/handle/123456789/27772 |
|
Language |
English
|
|
Relation |
Not Available;
|
|
Publisher |
Agriculture Research Communication centre
|
|