血浆及精清睾酮的酶免疫分析法研究

Studies on the Enzymeimmunoassay of Testosterone in Blood Plasma and Seminal Plasma

用混合酸酐法合成睾酮-3-(羧甲基)肟-过氧化物酶和睾酮-17-(羧甲基)肟-过氧化物酶两种结合物,并用睾酮抗血清和睾酮单克隆抗体建立鉴定了睾酮的两种固相微滴板酶免疫分析法(T-EIAa,T-EIAb),其中T-EIAa符合方法学鉴定指标.用T-EIAa测定种公牛血浆及精清睾酮含量,血浆样品经二氯甲烷抽提,而精清样品用直接法测定.T-EIAa测定血浆样品的结果与放射免疫测定密切相关(r=0.83,n=12),种公牛的血浆及精清睾酮含量在个体间存在显著差异,精液品质良好组的精清睾酮含量显著地高于较差组,而血浆睾酮含量差异不显著.

Two conjugates, testosterone-3- (carboxymethyl) oximeperoxidase and testosterone-17-( carboxymethyl ) oximeperoxidase, were synthetized by the mixed anhydride method. Using these conjugates and testosterone antiserum and testosterone monoclone antibody, two kinds of solid-phase microtitre plate enzymeimmunoassay of testosterone ( T-EIAa, T-EIAb ) were developed and appraised by methodology. The T-EIAa satisfied the normal criteria of methodology. The testosterone concentrations in blood plasma and seminal plasma in stud bulls were determined by the T-EIAa. The samples of blood plasma were extracted with dichloromethanc, while the samples of seminal plasma was measured directly. Results obtained by T-EIAa and radioimmunoassay were closely related in samples of blood plasma ( r = 3.8332, n=12) . The testosterone concentrations in blood plasma and seminal plasma varied considerably among the individuls in stud bulls. The testosterone concentrations in seminal plasma in the group with good semen quality was higher significantly than that in the group with bad semen quality. However, no statistical correlation was observed between testosterone concentrations in blood plasma and semen quality.