摘要
为可视化检测羊肉中鸭肉源成分,该研究将跨越式滚环等温扩增(Saltatory rolling circle amplification,SRCA)技术与荧光技术相结合,以生长激素(Growth hormone,GH)基因为靶基因分别设计并筛选羊、鸭特异性引物,拟建立可视化SRCA检测方法。对该方法进行特异性、灵敏度和检出限试验。结果显示:山羊和绵羊样品呈现绿色荧光,判定为阳性,13种非羊样品呈现橙黄色荧光,判定为阴性;鸭样品为阳性,14种非鸭样品均为阴性;表明该方法特异性良好。可视化SRCA分别检测羊、鸭DNA的灵敏度均为2.0×10^(1)fg/μL。可视化SRCA检测羊肉中鸭肉源成分的检出限为0.001%(w/w)。对29份市售羊肉样品中鸭肉源成分的检测得知,可视化SRCA法检出率为17.24%,行标NY/T 3309-2018中方法检出率为10.34%。该方法经济、简便,结果肉眼可见,更适用于条件相对落后的实验室或资源相对匮乏的基层检测机构。
In order to visually detect duck meat components in mutton,this research combines the saltatory rolling circle amplification(SRCA) technology with the fluorescence technology.To establish the visual SRCA detection method,the specific primers for goat and duck were designed and screened,respectively,with the growth hormone(GH) gene as the target gene.The results suggested that the goat and sheep samples emerged green fluorescence,which were judged as positive results.And the 13 nongoat samples presented orange fluorescence,which were judged as negative results.The duck sample was positive,and the 14 non-duck samples were negative.Therefore,the specificity of the detection method was excellent.The sensitivities of visual SRCA for detecting DNA of goat and duck were both 2.0×10^(1) fg/μL.The detection limit of visual SRCA for detecting duck meat components in mutton was 0.001%(w/w).The detection rate of duck meat components using visual SRCA in 29 commercially available mutton samples was 17.24%,while the detecton rate of the method in industry standard “NY/T 3309-2018” was 10.34%.The results are visible to the naked eye,which is economic,simple and convenient.The visual SRCA method is suitable for laboratories with backward conditions or basic testing institutions with relatively scarce resources.
作者
李韦霖
郭威
梁梓薇
孙旭飞
刘杰
杨倩
范少华
张伟
LI Weilin;GUOWei;LIANG Ziwei;SUN Xufei;LIU Jie;YANG Qian;FAN Shaohua;HANG Wei(College of Food Science and Technology Hebei Agricultural University,Boing 071000,China;College of Science and Technology,Hebei Agricultural University,Cangzhou 061100,China;College of Public Health,Hebei University,Baoding 071002 China;Hebei Software Institute,Baoding 071030,China;College of Life Sciences,Hebei Agricultural University,Baoding 071000,China;Hebei Key Laboratory of Analysis and Control of Zoonotic Pathogenic Microorganism,Baoding 071001,China)
出处
《食品科技》
CAS
北大核心
2022年第3期311-317,共7页
Food Science and Technology
基金
国家自然科学基金项目(31371772)
河北省自然科学基金重点项目(C2019204342)
河北省重点研究开发计划项目(18275501D)
中央引导地方科技发展资金项目-基础研究项目(216Z5501G)
河北农业大学食品加工学科群经费项目(2021-06)
河北省在读研究生创新能力培养项目(CXZZBS2019091)
博士后科研项目择优项目(B2021005007)。
