免疫磁珠结合PCR-试纸条法鉴定金枪鱼成分的研究  

Identification of tuna ingredients by immunomagnetic beads combined with PCR-based lateral flow assay

作　　者：赵良娟 邢冉冉 庞璐 刘亚华 刘金玉 郑文杰 ZHAO Liang-Juan;XING Ran-Ran;PANG Lu;LIU Ya-Hua;LIU Jin-Yu;ZHENG Wen-Jie(Animal&Plant&Foodstuffs Inspection Center of Tianjin Customs District,Tianjin 300461,China;Chinese Academy of Inspection and Quarantine,Beijing 100176,China;Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin 300387,China)

机构地区：[1]天津海关动植物与食品检测中心,天津300461 [2]中国检验检疫科学研究院,北京100176 [3]天津师范大学,天津市动植物抗性重点实验室,天津300387

出　　处：《食品安全质量检测学报》2021年第13期5260-5265,共6页Journal of Food Safety and Quality

基　　金：国家重点研发计划项目(2017YFF0211301);海关总署科技计划项目(2019HK096)。

摘　　要：目的建立一种免疫磁珠结合PCR-免疫金标试纸条快速检测金枪鱼成分的方法。方法使用免疫磁珠法提取金枪鱼样品的核酸,针对金枪鱼基因组特异性基因片段设计引物,优化反应体系和反应条件,建立金枪鱼成分PCR产物试纸条检测方法,验证方法的特异性、灵敏度和稳定性,并对市售金枪鱼产品进行检测。结果免疫磁珠法提取市售金枪鱼样品DNA的纯度高。所建立的PCR-试纸条方法特异性强,仅金枪鱼能扩增出229 bp特异性DNA片段,试纸条检测线变红,其他鱼类均无非特异性扩增条带,试纸条检测线未变红色;灵敏度高,达到0.01%;方法快速,PCR产物的试纸条判读时间5 min。结论该方法具有特异性强、灵敏度高、稳定性好、简便环保等优点,适用于金枪鱼成分的快速检测。Objective To establish a method for rapid detection of tuna ingredients by immunomagnetic beads combined with PCR-based lateral flow assay.Methods A nucleic acid of a tuna sample was extracted by using an immunomagnetic beads,a primer was designed aiming at a tuna genome specific gene fragment,a reaction system and reaction conditions were optimized,a tuna ingredients PCR-based lateral flow assay detection method was established,the specificity,sensitivity and stability of the method were verified,and a commercial tuna product was detected.Results The purity of DNA extracted from commercial tuna samples by immunomagnetic beads was high.The established PCR-based lateral flow assay method had strong specificity,only tuna samples could amplify 229 bp specific DNA fragments,and the detection line of the test strip turned red.There was no non-specific amplification band in other fish,and the detection line of the test strip did not turn red.The sensitivity was high,reaching 0.01%.The method was rapid,and the test strip interpretation time of PCR products was 5 min.Conclusion This method has the advantages of strong specificity,high sensitivity,good stability,simplicity and environmental protection,and is suitable for rapid detection of tuna ingredients.

关 键 词：免疫磁珠 金枪鱼成分 PCR-试纸条 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]