A sandwich enzyme linked immunosorbent assay (S-ELISA) for detection of MrNV in the giant freshwater prawn, Macrobrachium rosenbergii (de Man).
A sandwich enzyme-linked immunosorbent assay (S-ELISA) was developed to improve diagnosis of white tail disease of the giant freshwater prawn, Macrobrachium rosenbergii, caused by the nodavirus, MrNV. Polyclonal antibodies were produced by immunization of Balb/C mice using a purified suspension of the virus and IgG anti-MrNV were purified from ascitic fluid. A sandwich method was successfully developed, coating first with unlabelled antibody and detecting trapped antigens with a second biotinylated antibody. Reaction was demonstrated using an avidin-peroxidase conjugate. Tissue extracts from M. rosenbergii infected with MrNV or purified viral extracts (control) were successfully identified in an individual ELISA, thus confirming the validity of the method. This S-ELISA should be the technique of choice for epidemiological studies of this disease and is a rapid and inexpensive assay with high specificity and sensitivity.
Romestand
B
B
UMR 5098 DRIM, CNRS/IFREMER/UM2, Université Montpellier II, Montpellier Cedex 5, France. romestand@crit.univ-montp2.fr
Bonami
J R
JR
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D030381:Nodaviridae / Q000276:immunology* / Q000302:isolation & purification* / Q000648:ultrastructure; D033541:Palaemonidae / Q000821:virology*; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D007074:Immunoglobulin G