Establishment Of A Method For Identification Diseased Pork By POD And Hb As Two Targets

In the current methods of identifying diseased pork,sensory inspection and qualitative inspection cause experiential and subjective results and fail to provide accurate graphs and quantitative data,which is not conducive to the identification of pork carcass and results in potential risks to the safety of pork.To identify and diagnose diseased pork,this study established a dual detection targets with a peroxidase concentration and a residual Hb concentration of pork by hemoglobin-linked immunosorbent assay.The results of this study mainly focused on the following aspects.1.Preparation of porcine Hb monoclonal antibody and establishment of ELISA.BALB/c mice were immunized with porcine Hb,11 hybrid cell tumor strains were obtained after 4 times of subcloning and screening by hybridoma technique,which can stably secrete monoclonal antibodies with good specificity according to the result of western blot.In this study,Z4 was used as a coating antibody and G as the enzyme-labeled antibody to establish a porcine Hb double antibody sandwich ELISA assay according to the highest titer(6.4×10~5)of the four monoclonal antibodies(G,H,Z4,Z5).2.Comparative analysis of several diseased pork detection methods.The pH,POD,Hb was determined by determining the related indexes of diseased pork with different treatment methods and study the accuracy of the four methods.The results showed that the meat infusion under machine agitation method were better than those by hand shearing;The detection accuracy of POD detection method and hemoglobin ELISA detection method was higher than that of pH detection method and hemoglobin absorption value detection method.However,the single test method cannot be used as a criterion for identifying diseased pork.3.Establishment of a method of identification diseased pork by dual targets detection with POD and Hb.it is the for the meat samples to be soaked for 10 minutes and diluted10 times were the best pre-treatment condition.Through the exploration of the optimal detection conditions of POD and Hb on pork,a differential diagnosis method was established,in which the POD concentration and Hb residual concentration in pork were double detection targets,in which the identification accuracy can reach 95.5%.According to the statistics of compliance rate,the R value of diseased pork and healthy pork as a judging standard was established,that is,pork was healthy when R value was less than or equal to 300,and diseased pork was identified when R value was more than or equal to 400.In summary,this study established a dual detection targets with a peroxidase concentration and a residual Hb concentration of pork by hemoglobin-linked immunosorbent assay by a porcine hemoglobin ELISA method,which provided theoretical basis and technical support for the identification of clinical diseased pork.