摘要
以鲢鱼（Hypophthalmichthys molitrix）卵黄蛋白原（Vitellogenin,Vtg）为抗原,卵黄蛋白原抗血清为抗体,建立了检测鲢鱼体内卵黄蛋白原的间接酶联免疫吸附反应（ELISA）方法,组装卵黄蛋白原ELISA试剂盒,并对其性能进行鉴定。结果表明,建立的ELISA间接法检测浓度为6.1-396.0 ng/m L,灵敏度为6.1 ng/m L,且在该范围内标准曲线线性良好;稳定性试验结果表明,在系列抗原标准品中添加1%蔗糖、20%甘油和0.000 5%Mg Cl2时,试剂盒在37℃条件下放置7 d,其标准曲线灵敏度和线性基本保持不变,说明筛选的该稳定剂配方对ELISA试剂具有良好的保护作用。
A blocking ELISA kit was established for detecting Vtg in the silver carp and its properties were examined. The technique was developed with antiserum against vitellogenin （Hypophthalmichthys molitrix）（Vtg） as antibody and Vtg as antigen. The results showed that the working range of the Vtg-kit was 6.1 to 396 ng/mL,the sensitivity was 6.1 ng/mL,and the standard curve showed a good linearity in working range. Stability experiment showed that the sensitivity and line performance of kit remained unchanged under 37 ℃ for 7 days,indicating that the selected stabilizer had a signification protective effect on the ELISA kit.
出处
《湖北农业科学》
2016年第13期3490-3493,3497,共5页
Hubei Agricultural Sciences
基金
武汉市国际科技合作计划项目(2014030709020314)
湖北省科技支撑计划项目(2014BBA163)
湖北省重大科技创新计划(2015ABA038)
湖北省农业科技创新中心项目(2016-620-000-001-036)
