Early Pregnancy Diagnosis With Donkeys Serum PMSG Immunology

With the large-scale and intensive development of donkey breeding,the breeding efficiency has become an important factor restricting the breeding efficiency,through early pregnancy diagnosis,reducing empty pregnancy is an important measure to improve reproductive efficiency.but the early pregnancy diagnosis of donkey mainly relies on"rectal palpation method".At present,the diagnosis of pregnancy in donkeys mainly relies on"rectal palpation method",and the diagnosis time is about 60 days after mating,which not only delays the pregnancy test and prolongs the empty pregnancy period,but also greatly increases the risk of transmission of zoonoses.In this study,a mature PMSG test method was established through three parts,namely,the immunological test condition of PMSG in pregnant horses,the best detection period of PMSG after mating and the immunological test compliance rate of PMSG,which laid a foundation for the preparation of early pregnancy test strips in donkeys.The research results are as follows:Establishing PMSG immunological detection method.The purity of commercial PMSG antigen was detected by SDS-PAGE.The Dot blot experiment verified that PMSG can bind to the antibody.Western Blot verified that the binding of PMSG antibody to antigen by spatial epitope,and the PMSG concentration and absorbance value standard curve line was further established by ELISA.The fitting equation was y=0.0045x+0.0202,R²=0.998;Secretion regulation of PMSG in female donkey after pregnancy.The serum samples of donkeys at 2,3,4,5,6,7 and 8 weeks after breeding were collected,and the PMSG content in serum was detected by ELISA.Serum PMSG can be detected from the 5th week and the content is gradually increased.And the concentration reached a peak of 80 IU·mL^-1 at the 50th day,then declined subsequently.The PMSG in serum has not been detected during 2-8 weeks for unpregnant donkeys;Clinical validation.Choosing fifteen breeding female donkeys and 15 head pregnant donkeys were examined by rectal examination,established by the test of the serum immunology testing methods 5 weeks stepmother donkey serum mating,found that pregnancy 7 head donkey,not pregnancy 8 head donkey,positive clinical pregnant coincidence rate was 46.7%,the detection of breeding donkey serum six weeks stepmother,found that at 14 head donkey,not pregnancy 1 head donkey,donkey positive pregnancy clinical coincidence rate was 93.3%,the detection of breeding donkey serum seven weeks stepmother,found that at 15 head donkey,not pregnancy 0 head donkey,donkey positivepregnancy clinical coincidence rate was 100%.The results of serum immunological test showed that there were 15 negative controls and the clinical coincidence rate was 100%.In summary,the period to detect early pregnancy female donkeys by serum PMSG enzyme-linked immunosorbent assay was 5 weeks after breeding,and the best detection period was 6 or 7 weeks after breeding.