摘要
本实验利用自建的奶牛乳汁孕酮竞争ELISA法（酶标抗原法）研发了奶牛乳汁孕酮ELISA试剂盒。通过对试剂盒的特异性、灵敏性、重复性、稳定性和回收率等主要性能进行优化试验。结果表明：试剂盒可以在4℃条件下保存9个月,与睾酮、雌三醇、雌二醇无特异性反应,灵敏度为0.14 ng/mL,批内差异为4.11%,批间差异为6.26%,回收率为93.80%~107.60%,变异系数为5.65%。应用该试剂盒检测乳汁孕酮诊断奶牛妊娠与未妊娠的准确率分别为93.3%和95.8%。因此,该试剂盒性能良好,妊娠诊断效果理想,为今后在奶牛生产中的广泛应用奠定基础。
In this study, a milk progesterone ELISA detection kit for dairy cows was developed using enzyme labeled antigen method established in this laboratory. Main performances of ELISA kit were optimized by determining specificity, sensitivity, repeatability, stability, and recovery for ELISA kit. Results showed that the ELISA kit could store for 9 months at 4℃, there was no cross-reaction between progesterone and testosterone, estradiol, estriol, the sensitivity was 0.14 ng/mL, intra-assay variation was 4.11%, inter-assay variation was 6.26%, the recovery rate was 93.80%-107.60%, coefficient of variation was 5.65%. According to milk progesterone levels, the diagnosis accuracy of the ELISA kit was 93.3% for pregnancy and 95.8% for non-pregnancy, respectively. Therefore, the developed ELISA kit has good performance for diagnosis of pregnancy in dairy cows. It will provide a basis for a wide application in dairy cows production.
出处
《中国畜牧杂志》
CAS
北大核心
2015年第21期54-58,共5页
Chinese Journal of Animal Science
基金
黑龙江省攻关项目(GB07B205)
黑龙江省农垦总局攻关项目(HNK11A-08-01-09)
