Research On Several Methods For Detecting Non-avian Biological Products Of Exogenous Virus

Veterinary Pharmacopoeia of the People’s Republic of China (2010Edition) was formulated in thetesting of non-avian exogenous virus. Tests are normally done with three methods: cytopathic effectdetection, hemadsorption test and fluorescence antibody detection, to confirm whether they are pollutedby exogenous virus after sample has been cultured by the selected cell culture for14days. Phenomenasuch as cell lesion, RBC adsorption or the specific indicate exogenous contaminated thereforedisqualification. This experiment is to improve the accuracy of the experiment, define contaminated virusspecies based on the proven unqualified products, and further realize the targeted prevention.In the cytopathic effect(CPE) experiment,we culture and observe6kinds of the production cellsinoculated with8kinds of production or inspection required virus, the CPE experiment results except forjudging products to be unqualified(pharmacopoeia rules),it also can be determined that the virus whichmake Vero appear CPE include PPRV,PRV,GTPV and Canine parainfluenza virus; producing CPE in PK15include PRV,PPV, in Sheep testicular cells appear CPE have PPRV,PRV,GTPV; appearing CPE in ST cellshas PRV; generating CPE in BHK21include PRV and FMDV;CSFV cells don’t produce the CPE.Hemadsorption experiment is to detect the virus which make cells have erythrocyte adsorption action.We chose6kinds of virus to vaccinate in4kinds of cells respectively and then pick out the virussusceptible cells. The result showed that PPV can make PK15have erythrocyte adsorption ability, makeSheep testicular cells have erythrocyte adsorption ability is GPV, CPIV can make Vero have erythrocyteadsorption ability in Vero cells.Fluorescent antibody test is a method using known antibody test samples to detect whether there arecorresponding antigens by fluorescence method. The regulated dilution ratio of primary and secondaryantibody by commercially available,some rations in the experiment often have no or poor fluorescenceeffect. In this study,to compare the fluorescence effect influence of different cell concentration,first andsecondary antibody concentration, then finds out the optimal proportion: the fluorescence slice had the bestobservation effect when PPV stain the first antibody titers ranged from1:800to1:900, and secondaryantibody titers were1:500; the fluorescence slice had the best observation effect when CSFV stain the firstantibody titers were1:4and secondary antibody titers were1:500; the fluorescence slice had the bestobservation effect when BVDV stain the first antibody titers ranged from1:150to1:200and secondaryantibody titers were1：200.