检测睾酮、孕酮、雌酮与雌三醇的双抗体酶免疫分析法  

DEVELOPMENT OF DOUBLE-ANTIBODY ELISAs FOR TESTOSTERONE, PROGESTERONE, ESTRONE AND ESTRIOL DETERMINATION

作　　者：郭大智[1] 

机构地区：[1]四川农业大学核技术农业应用研究室,四川雅安625014

出　　处：《核农学报》2002年第1期45-48,共4页Journal of Nuclear Agricultural Sciences

摘　　要：本文报道了自制的试剂建立了检测睾酮、孕酮、雌酮和雌三醇的双抗体酶免疫分析法。实验表明 ,这些测定具有很高灵敏度 (最小可测量为 0 1～ 1pg 孔 ) ,标准曲线范围为 0～ 40pg 孔 ;同相关类固醇激素 (孕酮、孕烯醇酮、睾酮、雄酮、雌酮、雌三醇和皮质酮等 )的交叉反应率一般在 0 1 %以下。测定的批内和批间变异系数分别在5 8%～ 7 2 %与 9 5%～ 1 0 5% (n =8)范围内。向血中添加激素标准品后的回收率在 94%～ 1 1 0 %范围内。用自制防腐剂处理后 ,液态抗体 (包括第二抗体 )、酶标记物和已包被第二抗体的滴定板可保持活性 5～ 1 2个月或更久 ( 4℃～ -2 0℃ )。此测定可节约激素特异抗体 4～ 1 0倍。初测人和动物的血、奶、尿样品表明所建立的 4种测定方法可分别用于人和动物体液中睾酮、孕酮。Sensitive ELISAs for testosterone, progesterone, estrone and estriol measurement using second antibody techniquqe were developed. The key reagents used in the assays including hormone specific antibodies, goat IgG anti rabbit IgG and enzyme steroid conjugates were self prepared. The detecting limits of these assays were 0 5～1 0pg/well; the calibration curves were sensitive in the range of 0～40pg/well.The intra and interassay coefficients of variation were in the range of 5 8%～7 2% and 9 5%～10 5%(n=8), respectively. The cross reactivities with progesterone, pregnenolone, testosterone, androsterone, estrone, estradiol, estriol and corticosterone were mostly below 0 1%. Recovery rate of the 4 steroids added to human serum ranged 94%～110%. The liquid antibodies(including second antibodies), enzyme conjugates and second antibody coated plates remained workable for 150 to 360 days or more (4～20℃) after treatment with the conservative self prepared. The quantity of the hormone specific antibody needed in the present assays was only 1/4 to 1/10 of the previous ELISA, in which the plate was coated with hormone specific antibody. Preliminary tests for the 4 steroids in sera, milk or urine in pregnant women, pigs, dairy cows, chickens and giant pandas showed that these ELISAs with high sensitivity, reproducibility, specificity and long storage life can be used for progesterone, testosterone, estrone and estriol determination in human and animal body liquid.

关 键 词：双抗体酶免疫分析 睾酮 孕酮 雌酮 雌三醇 人体 动物 检测 

分 类 号：S852.4[农业科学—基础兽医学]