Progesterone And Estrone Sulfate Collaurum Test Paper Preparation And Application In Early Pregnancy Detecyion Of Shennongjia Golden Snub-nosed Monkeys(Rhinopithecus Roxellana)

The golden snub-nosed monkey(Rhinopithecus roxellana)is one of unique and endangered species of China,and it had been classified as class I national key protection animal of China and endangered animals in the appendix I List of CITES(the Convention on International Trade in Endangered Species of Wild Fauna and Flora).Shennongjia is the east edge of habitat of golden snub-nosed monkeys.There are approximately 1200 golden snub-nosed monkeys in Shennongjia.Low fertility is one of the major factors which threats the survival of these animals.Therefore,it is significant to recover the population of this precious monkey specie.The purpose of this experiment is to establish methods of progesterone(P4)and estrone sulfate gold immunochromatography assay strip,and then use these strips to detect early progrance of Shennongjia golden snub-nosed monkey on time..The main results are as follows:1.The results showed two hybridoma cell strains secreting monoclonal antibody(McAb)against P4 were screened,named 2C5 and 5D2.Monoclonal antibody that prepared through ascites in mice was idengtified as lgG2b and the titer was 1:2*104,and the concentration was 3.54mg/ml after purification.The antibody has considerable good specificity,there was no cross-reactivity reaction with other similar objects.2.In this study,the detection line of progesterone colloidal gold test paper strip was 25ng/ml,and no cross-reactivity reaction with other similar objects,that is to say the strip has a good specificity.We used the strip to detect the urine of golden snub-nosed monkeys in Shennongjia,and the results drawn that the the accurate rate of strips was 17/18.3.Estrone sulfate hapten synthesized by estrone3-sulfate sodium salt and identified by mass spectrometry,was respectively coupled with BSA,KLH and OVA through the carbodiimide method to obtain the complete antigens.Identification of complete antigens by UV spectrum.Compared with its hapten and coupled protein the UV spectrum of complete antigens had a significant change.4.In this study,only one hybridoma clones were selected against estrone sulfate,and named 1D3.The ELISA titer of this McAb was 1:1600.The sensitivity of estrone sulfate colloidal gold test paper strip was 10ug/mL.