| With the development of the economy,the circulation of livestock is extremely frequent,which makes the spread of brucellosis among livestock more rapid.At present,brucellosis diagnosis,immunization,prevention and other aspects have also developed rapidly,but there is still a lack of practical diagnostic techniques for brucellosis during the excretion period,which causes great difficulties in effectively preventing and controlling brucellosis and purifying cattle.The practical diagnostic technical process of bovine brucellosis excretion period is particularly important.In this study,the red tiger plate agglutination test(RBT),microtube agglutination test(MAT),fluorescence polarization test(FPA)and hapten agar diffusion test(NH-GD)were used for the detection of brucellosis antibodies.The coincidence rate,sensitivity and specificity of FPA and MAT were compared with the MAT results.The NH-GD detection method was used for the practical diagnosis of brucellosis in the excretion period,and the positive samples were tested by AMOS-PCR.Through the analysis of the test results,different detection methods were selected to establish a practical diagnostic technical process for bovine brucellosis,which was used in the monitoring and purification of cattle brucellosis.The specific results are as follows:1.Study on the immunological diagnosis method of bovine brucellosis.The four brucellosis antibody detection methods of RBT,MAT,FPA and NH-GD were used to detect 106 serum samples.The positive rate of RBT was 30.19%(32/106),the positive rate of MAT was 24.53%(26/106),and the positive rate of FPA was 30.19%(32/106).The positive rate was 18.86%(20/106),and the positive rate of NH-GD was12.26%(13/106).The differential diagnosis of NH-GD was positive in 13 cases,one was bovine type I and one was bovine type III by AMOS-PCR.2.Establishment of technical process for differential diagnosis of bovine brucellosis.Through the selection and combination of different detection methods for brucellosis,two technical processes for differential diagnosis of bovine brucellosis are established.Process 1: RBT primary screening,MAT diagnosis,NH-GD differential diagnosis,and AMOS-PCR typing;Process 2: RBT initial screening,FPA review,NH-GD differential diagnosis,AMOS-PCR typing.NH-GD positive cattle were culled and treated harmlessly.3.Application of practical diagnostic technology process in bovine brucellosis excretion period.In the application of process 1,3688 serum samples collected from cattle farm A were tested,and the infection positive rate was 0.57%(21/3 688).The positive samples were detected by AMOS-PCR and sequenced as bovine type I,A19 vaccine strain.The application of the second process,the detection of 2 600 bovine serum samples collected from the B cattle farm,the infection positive rate was 0.46%(12/2 600),the positive samples were detected by AMOS-PCR and sequenced as bovine type I,A19 vaccine strain.The infected cattle detected were culled and treated harmlessly.The two practical diagnostic technical procedures established in this study can distinguish vaccine immunization from naturally infected cattle,and can be used for brucellosis monitoring in cattle farms;PCR detection and sequencing showed that the infection was caused by A19 vaccine strain,because of large doses,repeated Caused by irregular immunization;it is recommended that cattle farms immediately cull and harmlessly treat NH-GD positive cattle;the purpose of controlling and purifying brucellosis is achieved by culling infected cattle and cattle farms. |
