Establishment Of Self-coating Radioimmunoassay Using Progesterone Mono-clonal Antibody And Its Application

Progesterone (P4) is a major steroid hormone which directly control the reproduction of female mammals. Analyzing results of progesterone from milk samples or blood samples can provide ovarian information, and make the monitoring of female mammal reproductive status possible. Potential uses of progesterone testing in reproductive management program are: I) identif~ying errors in detection of estrus; 2) predicting time of estrus; 3) non- pregnancy diagnosis in early stage; 4) differentiating types of ovarian cysts, and 5) evaluating response to endocrine therapy. Liquid-phase radioimmunoassay (3H-P4) and enzymeimmunoassay(EIA) are the techniques used for detecting animal reproductive performance by analyzing progesterone in blood and milk in our country. However, the former would produce radioactive wastes, which need special treatment for disposal, and the results got from IEIA sometime are not very precise. The technique of Self-coating solid-phase radioimmunoassay (Sc- RTA) of progesterone is developed with progesterone monoclonal antibody. The sensitivity of the technique is 0.O6nmoIIL. The intra- and inter-assay coefficient of variation is 6.8% and 9.3%, and the average recovery is up to 95.73%. There is a very good correlation between Sc-RIA and DPC. 3H-P4 and ??-P4 were incorporated with dry filter paper, and 90% ethanol was used to extract P4 in the solid sample-filter paper. The extraction efficiency is 95.01 ?.80% and 94.09?.03% respectively. According to the 損ower- point system?suggested by Cerceo, the score of Sc-RIA is 31 points (a method scored higher than 28 points is considered as acceptable), which confirms the reliability of Sc-RJA method developed in this study. By using the Sc-RIA, estrous cycle, pregnancy and abnormal luteal 53 function were studied. The results show that the changing profiles of P4 concentration in skim milk and filter paper samples were similar. In the estrous cycle of normal cow, the correlation coefficient of P4 concentration in skim milk and filter paper samples is 0.8792 ?0.070 and the estrous cycle interval is the same, both of them are 22 ?2.1 8d. Foot P4 concentration in filter paper and skim milk samples is 1.6481?0.6O23nmoIIL and 0.3184 ? 0.2549nmo1[L, respectively. The changing profiles of P4 concentrations in skim milk and filter paper samples of artificial inseminated cows is similar and the correlation coefficient is 0.9178?.0053. The Sc-RLA can also be used in early pregnancy diagnosis for milk cow. Based on progesterone Sc-RIA of skim milk and filter paper samples, the results of pregnancy diagnosis in early stage show that the accuracy of pregnant and non-pregnant is 89.47% and 100%; 87.6 1% and 95.47%. respectively. The ovarian information through milk or blood progesterone Sc-RIA makes the monitoring of the female mammal reproductive status possible.