Development Of Indirect Dot-ELISA Assay For The Detection Of Clostridium Perfringen Of Type D From Bovine Antibody

Cl.perfringens that was widespread in the natural world can cause enterotoxaemia of calf even death under certain condition.So there is no integrity serology detection method was used by detecting specific antibody that produce by Cl.perfringens.The study purpose was to found a kind of quickly,exact and aeschynomenous serological diagnosis by depurated tropina.Base on Clostridium perfringens of purified antigen was produced from type strain of D C60 by supersonic wave,SephadexG-200,SDS-PAGE and Western blot analysis.The result showed that purified antigen produced satisfactory specific immune response with antibody from Bovine blood serums.The result shown optimal concentration of antigen was 1.95μg/mL; optimal concentration of serum dilution was1:320;optimal condition of HRP-labeled rabbit anti-bovine IgG was 1:2 000;the level of spalit antigenic was 40.6μg/mL when the same serum sample that was 1:320 was detected by indirect Dot-ELISA and purification antigen was 1.95μg/mL;the positive of 100 samples rate was highest from Dunhua;the detection result of antibody average titre of 30 positive samples was 1:736 and 1:17 by Dot-ELISA and AGID;the relative sensitivity indirect Dot-ELISA was 43 time of AGID;Cross reaction showed indirectDot-ELISA assay to have higher specificness.Indirect Dot-ELISA that detected antibody of Cl.perfringens of type D was to found by using purification antigen;there were fortes not only high degree of specificity and high sensitivity of ELISA but also fewer antigen usage amount,quick and convenient about indirect Dot-ELISA,the diagnosis and prophylaxis of bovine Clostridium perfringens of type D was provided dependable experiment method.