Development Of McAb To GPMV And The Study On ICS To Diagnose GPMV

In this study , We establish a rapid and sensitive sandwich immunochromatographic strip(ICS) for goose paramyxovirus (GPMV) detection.Part one is get the monoclonal antibodies(McAbs) to GPMV. JG04 strain virulent GPMV was purified by PEG6 000 followed by Sephdex G200 column chromatography,protend ,viral protein 1.283mg/mL,HA 2^9～10.Immuning BALBC/C mouse and coating ELISA with antigen that is purified GPMV . Funsion by PEG1 500, the fusing rate is 46.5%.;Our screening of the hybridoma cells by indirect ELISA and HI,the positive rate is 78.4%; Positive hybridoma cell were subcloned three times by limited dilition.Two hybridoma cell lines which can stably secrete McAb was obtained and designated 3A₅ and 2C₆. And the ELISA titers of 3A₅ in cell supernatant and Ascitic were 1:51 200 andl:512,none HI titers; the ELISA titers of 2C₆ in cell supernatant and Ascitic werel:6 400andl:64, HI titers were 2⁴ and 2⁷.Part two is develop a rapid and sensitive sandwich immunochromatographic strip. Tri- sodium citrate with aqueous gold chloride were warmed up and mixed to make gold sol; The chicken IgG(anti-JG04 strain GPMV) was immobilized to a defined detection zone(T) on a porous nitrocellulose membrane, while 3A₅ was conjugated to colloidal gold which served as a detection reagent.The GPMV-containing allantoic fluid sample was added to the membrane and allowed to react with 3A5-coated particles. The mixture was then passed along the porous membrane by capillary action past the chicken IgG in the detection zone, which will bind the particles that had GPMV bound to their surface, giving a red color within this detection zone with an intensity proportional to GPMV concentration. In the absence of GPMV, no immunogold was bound,to the solid-phase chicken IgG.It was found that the GICA testing strip showed no cross-reactions with the allantoic fluid of A1V, EDSV, IBDV, IBV, GPV , CELO and the virus antigen of MV and CDV , to show a specific reaction , reacted with the allantoic fluid of ND viruses of different origin in goose, peafowl , pigeon , Lasota strain and chickens, to show a extensive detection of different origin-ND viruses.This immunochromatographic strip is based on an immunochromatographic procedure that utilizes...