摘要
为了完善已建立的孕酮免疫生物传感器的检测性能,本试验应用碳化二亚胺法将11α-羟基孕酮半琥珀酸和OVA制备成孕酮完全抗原,其浓度达0.6 mg/m L。在此基础上建立了以乳汁为基质的孕酮生物传感器的标准曲线(y=-4.9461x+8.7982,R2=0.993 2),进一步确立了检测乳汁孕酮的线性范围为0.31 ng/m L^50ng/m L,检测限为0.31 ng/m L,灵敏度为0.15 ng/m L,而且孕酮与皮质酮、皮质醇、雄烯二酮的免疫交叉反应较低,表明特异性较好。批内、批间变异系数分别为7.7%和7.5%,回收率为95.8%~115.7%,生物传感器稳定性在4℃保存4 d。该优化的奶牛乳汁孕酮免疫生物传感器的性能参数符合规定的标准,为检测妊娠、乏情和繁殖障碍疾病的奶牛乳汁孕酮含量提供了工具。
The aim of this study is to improve the established immune biosensor. The progesterone complete antigen was prepared using 11 α-hydroxy-progesterone hemisuccinate and OVA by carbodiimide method, and its concentration was up to 0. 6mg/mL. Then the progester- one biosensor standard curve was set up （y = -4.9461x＋8. 7982, R^2 =0. 9932）. The linear range of detection was 0. 31-50ng/mL, the lim- it of detection was 0. 31ng/mL, and the sensitivity was 0. 15ng/ml. Immunologic cross-reactivity was very low between progesterone and corticosterone, cortisol, and rostenedione. Intra-assay and inter-assay coefficient of variation were 7.7% and 7. 5 %, respectively; the recovery rate was between 95.8% and 115.7%. The optimized immune biosensor parameters could meet the prescribed standard. This study provides a useful tool for detection of milk progesterone level in dairy cows with pregnancy, estrus and ovary disorders.
出处
《畜牧与兽医》
北大核心
2015年第9期10-14,共5页
Animal Husbandry & Veterinary Medicine
基金
黑龙江省科技厅攻关项目(GB07B205)
黑龙江省农垦总局科技攻关项目(HNK11A-08-01-09和HNKXIV-08-01a)
关键词
乳汁
孕酮
免疫生物传感器
优化
milk
progesterone
immune biosensors
optimization
