Preliminary Establishment Of An Indirect ELISA Method For Sika Deer Mycobacterial Disease

Tuberculosis is a chronic wasting infectious disease.It is a zoonotic disease caused by Mycobacterium tuberculosis(MTB)invading humans and animals.Tuberculosis is spread mainly through feces and saliva.In recent years,it has spread widely in red deer and sika deer.The main pathogen of deer tuberculosis is Mycobacterium bovis.The widespread spread of the disease is a serious hazard to the development of the deer industry.In order to urgently find a method for rapid diagnosis of sika deer tuberculosis,this study used the immunogenicity of the 38 k Da protein secreted by Mycobacterium tuberculosis as a coating antigen.To establish an indirect ELISA method to detect deer tuberculosis.In the experiment,sika deer serum Ig G protein was used to immunize healthy male rabbits to prepare rabbit anti-sika deer polyclonal antibodies.The 38 k Da gene was combined into a 38 k Da gene by referring to the complete gene of Mycobacterium tuberculosis strain H37 Rv,and the 38 k Da gene was connected to the expression vector,and the 38 k Da protein was induced to express and purified in Escherichia coli.At the same time,an HRP-labeled rabbit anti-deer Ig G polyclonal antibody was prepared,and an indirect ELISA diagnostic method was initially established for the diagnosis of sika deer tuberculosis.In this experiment,rabbit anti-sika deer Ig G antibody and 38 k Da protein were successfully prepared,and an indirect ELISA detection method was established.The results show that the 38 k Da protein is highly immunogenic.The indirect ELISA method was used to preliminarily verify the 20 negative sera and 10 positive sera detected in the tuberculin test,and the test results were 100% consistent,indicating that the indirect ELISA method is feasible for detecting deer tuberculosis.It provides a new way for the rapid diagnosis of sika deer tuberculosis and the development of therapeutic drugs.