단일클론항체를 이용한 Milk Progesterone Test(EIA) 측정법의 개발과 이에 의한 소의 발정 및 임신조기진단의 정확도 향상에 관한 연구

Development of Milk Progesterone Test(EIA) using Monoclonal Antibody and It's Application to Estrus-and Early Pregnancy Detection in Dairy Cattle

A simple and sensitive microplate enzyme immunoassay(ELISA) was developed for progesterone, based on progesterone monoclonal antibody as anti-progesterone, horseradish peroxise(HRP) as enzyme-label and tetramethylbenzidine(TMB) as substrate. The assay has a sensitivity of 5pg-120pg/well and intra- and inter assay coefficients of variation for progesterone standard curve(0.1ng-3.2ng/ml) were ranged 4.4-10.6% and 5.-12.6%, respectively. They assay is performed in less than two hours and provide reliable values to differentiate among samples from day 0(A.I.), day 14 and day 19. The discriminatory levels for early pregnancy diagnosis are [>10ng(day 19) & decreasing rate <1.5 : pregnancy] and [ 7ng & decreasing rate 1.5 : non-pregnancy]. The accuracy of the pregnancy diagnosis for cows classified as positive(pregnancy) and negative(non-pregnancy) were 96% and 100%, respectively.