摘要
为检测食品中睾酮的残留,合成睾酮人工抗原并建立一种用于检测睾酮的酶联免疫方法(ELISA)。通过混合酸酐法将肟化的睾酮与鸡卵清蛋白偶联,合成人工包被抗原,与睾酮单克隆抗体竞争结合,根据光密度(OD)值对睾酮定量。结果表明,经紫外吸收、红外光谱、SDS PAGE电泳验证人工抗原合成成功,建立的间接竞争酶联免疫标准曲线线性范围为1~25μg/L,最低检测限为0.2617μg/L。对该检测方法进行稳定性测定,将储存3、7、10、14 d的试剂盒进行标准品测定,所得批间差异均小于5%。同时进行样品加标回收试验,平均加标回收率在87.51%~102.83%,说明建立的睾酮ELISA检测具有很高的准确度,可以应用于实际样品中睾酮的检测。本研究为进一步开发睾酮酶联免疫检测试剂盒检测食品中睾酮残留奠定基础,为保障我国肉制品质量安全提供一种有效的技术手段。
To detect testosterone residues in food,testosterone artificial antigens were synthesized to establish enzyme-linked immunoassay(ELISA)for the detection of testosterone.The oximized testosterone was coupled with chicken ovalbumin by the mixed anhydrides assay to synthesize the artificial coating antigen,which competed with the monoclonal antibody of testosterone.And the quantification of testosterone was based on optical density(OD)values.The results show that the synthesis of artificial antigen was verified by ultraviolet absorption,infrared spectrum and SDS-PAGE electrophoresis.The dynamic range of the established indirect competitive ELISA standard curve was 1-25μg/L,with the minimum detection limit of 0.2617μg/L.The stability of the detection method was determined,and the kits stored for 3,7,10 and 14 days were tested for standard substances,and the difference between batches was less than 5%.Meanwhile,the recovery test of spiked samples was carried out,and the average recovery of spiked samples was between 87.51%and 102.83%,which indicated that the established testosterone ELISA assay had a high accuracy and could be applied to the detection of testosterone in real samples.Therefore,this research put a potential for the further development of ELISA kit for the detection of testosterone residues in food,and provide an effective technical means for ensuring the quality and safety of meat products in China.
作者
高璐
孙嘉笛
王利平
纪剑
孙秀兰
GAO Lu;SUN Jiadi;WANG Liping;JI Jian;SUN Xiulan(State Key Laboratory of Food Science and Technology,School of Food Science,Jiangnan University,Wuxi 214122,China)
出处
《生物加工过程》
CAS
2020年第4期452-456,共5页
Chinese Journal of Bioprocess Engineering
基金
国家重点研发计划(2018YFC1602802)。
关键词
睾酮
人工抗原
紫外吸收
酶联免疫方法
检测
肉制品
食品安全
testosterone
artificial antigen
UV absorption
enzyme-linked immunosorbent assay
detection
meat products
food safety
