Preparation And Identification Of Monoclonal Antibody For The Early Diagnosis Of Trichinellosis

Trichinellosis is a global zoonotic parasitic disease caused by trichinella spiralis. It is still prevalence in China. More than 150 mammals could be infected besides human. Trichinellosis could not only cost great expenses in animal industry but also threaten people's health. The trichinellosis has no typical clinical symptom, the clinical diagnosis is difficult, it is easy to confuse with other infection. Although muscle biopsy may diagnose, but often not easy to discover in the mild infection.The biopsy positive rate also only then about 50%, and it is not accepted easily by the people because of the organization limited influence. Therefore, seeks one specificity, the sensitive high diagnosis method to prevente and controll trichinellosis has the significance. ELISA is the present most sensitive method, its sensitivity may examine to each gram meat 0.01 tenia. The sensitivity may guarantee the security completely of meats. However experiment indicated that, the anti-trichinella spiralis immune body after to infect 2 weeks only then to be possible to found for diagnoses in the antigen examination blood serum with the trichinella spiralis myo-larva ES antigen, but circulates antigen (CAg) is the excretion initial secretes the (ES) antigen to be possible to examine in early days. Therefore, examines CAg is a very effective method to solve the trichinellosis early time to diagnose. Through the experiment, we discovered that antibody can be examined in 9th days.Therefore, the purpose of this experiment is preparing anti-trichinella spiralis newborn larva ES McAb, diagnosis kits for the establishment examination trichinella spiralis CAg early time diagnosis trichinella spiralis and provide the ideal experimental material.This experiment mainly includes three parts: 1. artificial infection trichinella spiralis Kunming mouse serum antibody dynamic change research: This experiment uses trichinella spiralis myo-larva excretion/secretion antigen (MES), newborn larva excretion/secretion antigen (NES) and the newborn larva and the adult antigen are the diagnosis antigen, examine the different number of days artificial infection trichinella spiralis Kunming mouse antibody. The result dedicated that newborn larva excretion /secretion antigen (NES) may react with the serum most early the antibody. Therefore, we selected the trichinella spiralis newborn larva ES antigen to prepare the monoclonal antibody. 2. Anti-trichinella spiralis newborn larva ES monoclonal antibody preparation: Infects the Balb/c mouse with the trichinella spiralis myo-larva, then killed the trichinella using Uygur rhzomorph after 14 days, the Balb/c mouse were infected once more after seven days, the spleen cell and the SP2/0 be fused. Obtain three high specificities, the high titer anti-trichinella spiralis newborn larva ES McAb hybridoma cell line, the names are TS-C6G7, TS-H6C3 and TS-C4F3. It not have the cross response with other parasite antigen. 3. anti-trichinella spiralis newborn larva ES monoclonal antibody immunology localization research: Kunming mouse were infected through intravenous for establishing animal model, then killed one mouse in 8, 10, 17, 20, 24, 28, 40th day, makes diaphragm 4μm the thick paraffin section, carries on the immunity fluorescence dyeing separately to three monoclonal antibodies. Dyes through the immunity fluorescence, discovered three monoclonal antibodies may use in trichinellosis early diagnosis. 4. Trichinellosis early diagnoses method: coating condition, the coating density, the monoclonal antibody best response density, the reaction time and the substrate reaction time were optimized, the trichinelosis early diagnosis method was established. It will be advantageous for the application.Conclusion: 1. trichinella spiralis newborn larva ES antigen can be used as the diagnosis antigen for early determinate the anti-trichinella spiralis antibody 2. Monoclonal antibodies were prepared for the early diagnosis of trichinellosis. 3. Monoclonal antibodies can be used for the early diagnosis of trichinellosis.4. The sandwich ELISA was estabilished for the early diagnosis of trichinellosis.