Establishment And Assessment Of Buffalo Tuberculosis Detection Methods

China has abundant resources of buffalo(Bubalus bubalus,Bb),and so as the cows,buffalos are now under the threat of Bovine Tuberculosis.(b TB)The b TB infected buffalos are also important reservoir of human TB.To control the disease,an accurate detection method is the key point.The detection methods we are using now are following cows' detection method and criteria.Many differences existed between buffalo and cow as they belong to the same family but different genus even they have a reproductive isolation.The results are as followed.Preparation of the standard positive samplesFirstly,we confirmed the buffalos we chosen were TB negative using skin test and antibody detection before BCG immunization.The sample was predicated as negative if the two methods were both negative.Then we give them a BCG immunization to mimic TB infection,check the antibody and skin thickness at particular point.This time the sample was predicated as positive if any of the two methods is positive.And finally,we had 71 TB negative samples and 45 positive samples after BCG immunization.Standardization of tuberculin skin testAnalyze the skin test result of buffalo and cow.Firstly,confirmed the significant difference of thickfold thickness between buffalo and cow.And then proved there were no significant difference among the kinds.Using the result of skin test during particular point then find out the variation from the changing of the criteria.At last,the cut-off value was confirmed as 2.495 mm by ROC curve.Preparation of Mc Ab and Pc Ab to Bb IFN-?In this study,we prepared monoclonal antibody and polyclonal antibody using mice and rabbits to Bb IFN-? that prokaryotic expressed.And 6 monoclonal cells with high titer named 2A9,7C4,9B6,9H2,10E2,10H4 were obtained.After purifying,we got monoclonal antibody Ig M.In addition,we immunized 3 rabbits to obtain hyper-immune serum.The highest serum titer belongs to the NO.3 rabbit.In conclusion,appropriate criteria for buffalo tuberculin skin test was determinated in the study.And monoclonal antibody and polyclonal antibody to Bb IFN-? were prepared to establish the ELISA detection nethod.All these together laid the foundation of the development of Bb IFN-? so as to the integrated control.