动物源食品中孕酮激素残留的时间分辨荧光免疫分析法的建立  

Establishment of Time-resolved Fluorescence Immunoassay for Progesterone Residues in Animal-derived Foods

作　　者：刘波[1] 华彦涛 马楠楠 赵炫 陈丽楠 袁利鹏[1] LIU Bo;HUA Yantao;MA Nannan;ZHAO Xuan;CHEN Li;nan;YUAN Lipeng(Guangdong Aib Polytechnic,Institute of Tropical Agriculture and Forestry,Guangzhou 511365,China;Guangzhou Wanlian Biotechnology Co.Ltd.,Guangzhou 510663,China)

机构地区：[1]广东农工商职业技术学院热带农林学院,广东广州511365 [2]广州万联生物科技有限公司,广东广州510663

出　　处：《现代食品科技》2024年第6期279-284,共6页Modern Food Science and Technology

基　　金：广东省教育厅工程技术研究中心项目(2019GGCZX010);广东省教育厅高职领军人才项目(食品加工技术专业)[20181423];广州市增城高层次人才项目[人才办201807];广东省普通高校青年创新人才类项目(2022KQNCX211);广东省攀登计划项目(pdjh2022b0914);广东农工商职业技术学院重点项目(xyzd1606)。

摘　　要：该研究以孕酮为研究对象,设计并制备了新抗原,通过动物免疫得到特异性兔抗体,基于所得抗原和抗体,进一步优化了抗原抗体浓度、反应缓冲液种类及其pH值等关键参数,最终在国内首次建立了用于动物性食品中性激素孕酮残留快速检测时间分辨荧光免疫分析法(TRFIA)。该研究确定最优实验条件为:抗原0.30μg/mL、铕标抗体0.40μg/mL、pH值7.2的TBST工作缓冲液,并建立标准曲线,方法的线性范围为0.46~6.94μg/L,IC_(50)达1.79μg/L,与睾酮交叉反应率为1.25%,与雌二醇、雌三醇等激素无交叉反应。实际样品的平均添加回收率为82.0%~113.0%,批内批间变异系数<5%,与高效液相色谱法(HPLC)检测结果的相关性良好(r2=0.988)。结果表明该研究所建立的TRFIA检测方法准确可靠、特异性好、灵敏度高,完全可用于动物源食品中孕酮残留的快速检测。In this study,progesterone was taken as the research object,a new antigen was designed and prepared,and specific rabbit antibody was obtained through animal immunization.Based on the obtained antigen and antibody,key parameters,such as the concentration of antigen and antibody,the type of reaction buffer and its pH value,were further optimized.Finally,a time-resolved fluorescence Immunoassay(TRFIA)for rapid detection of progesterone residues in animal-derived foods was established for the first time in China.In this study,the optimal experimental conditions were determined as follows:the concentration of antigen was 0.30μg/mL,the concentration of europium labeled antibody was 0.40μg/mL,the pH of TBST working buffer was 7.2,and the standard curve was established.The linear range of the method was 0.46~6.94μg/L,the IC50 reached 1.79μg/L,the cross-reaction rate with testosterone was 1.25%,and there was no cross-reaction with estradiol,estriol and other hormones.The average spiking recovery rate of the actual samples was 82.0%~113.0%,and the coefficient of variation within and between batches was less than 5%.The correlation with the analysis results obtained by high-performance liquid chromatography was good(r2=0.988).The results indicate that the TRFIA detection method established in this study was accurate,reliable,specific,and sensitive,and can be used for the rapid detection of progesterone residues in animal-derived foods.

关 键 词：动物性食品 孕酮 抗体 时间分辨荧光免疫分析 检测 

分 类 号：S859.84[农业科学—临床兽医学]