Identificaion Of The A Toxigenic Pasteurella Mutocida In Pigs With Atrophic Rhinitis And The Generation And Identification Of Monoclonal Antibodies

This study adopt separated using the bacterial separation and culturemethod, and then classified using biochemistry testing, 80 strains ofPasteurella mutocida(Pm) from 266 swabs of weaned piglet, collected fromthe five scale pig farms in Guangxi province. Nine strain toxigenicPasteurella mutocida were test by guinea pig skin necrosis experiment andmouse lethality assay, which were named to DNT⁺Pm-A,DNT⁺Pm-B,DNT⁺Pm-C,DNT⁺Pm-D,DNT⁺Pm-E,DNT⁺Pm-F,DNT⁺Pm-G,DNT⁺Pm-H,DNT⁺Pm-I. This nine DNT⁺Pm were confirmed that aredisease germ of AR, which by testing correspondence to nine swines' nasalsuffering PAR. DNT⁺Pm-C strain was tested DNT⁺Pm sero group A byImprovement Carter. DNT⁺Pm-C was confirmed to Pm by API autobiochemistry testing, which was sent to GuangXi Disease Testing Station.Using monoclonal antibody technique, with DNT⁺Pm-C afterweakens was immunized to BALB/C, fusion and screening by indirect ELISA, three monoclonal antibodies were generated, which weredesignated as 2D₁E₈,4A₁₁B₂,7E₃G₉. Antibody of ascitic fluid fromhybridomas were detected by indirect ELISA. The titers respectivelyranges 1:1600 to 1:12800. They can still stably excreted McAb after -80℃for three months.Three strain McAb were identificed by indirect immnofluorescentassay(IFA),Dot-ELISA and ACI-ELISA. 2D₁E₈,4A₁₁B₂,7E₃G₉ can reactewith DNT+Pm-C strain in 96 microplates using indirect immunofluorescentassay(IFA), especially 7E₃G₉ has strong fluorescent reaction, it has a titer of1:1280, which is highest. Using Dot—ELISA test antigen overlappingresponse, with three strain McAb and Staphylococcus aureus,Salmonella,Escherichia Coli,Streptococcus respectively; Simultaneouslywith three strain McAb and nine strain DNT⁺Pm respectively. Resultingthat three strain McAb can react with DNT⁺Pm, but can't react withStaphylococcus aureus,Salmonella,Escherichia Coli,Streptococcus. 7E₃G₉react Specificity was strongest. By three strain McAb (2D₁E₈,4A₁₁B₂,7E₃G₉)were boated in the 96 well microplates respectively, test DNT⁺Pm antigenfrom nine swines' nasal secretion. Coincidence rate is 100% compare toclinical diagnosis. Result shows that three strain McAb can recognizeDNT+Pm antigen from swines' nasal effectively. For three strain McAb areall react to DNT+Pm—C strain by SDS, indicating their antigen epitope of McAbs are linear epitope.. It can be apply to AR's rapid diagnosisreagent box.