摘要
根据GenBank中公布的猪线粒体色素细胞b基因序列设计6 条特异性环介导等温扩增(loop-mediated isothermal amplification,LAMP)引物,通过简化线粒体DNA提取步骤,优化反应体系,以常见的牛、羊、鸡、鸭、马、驴肉为阴性对照验证该方法的特异性,掺假率梯度稀释以验证该LAMP法的最低检测限,建立常见肉制品中猪肉成分的可视化LAMP检测方法。结果表明:该方法提取目的基因操作简单、稳定,该可视化LAMP检测法以4-(2-吡啶偶氮)-间苯二酚钠盐为指示剂,能准确检测出常见肉类中猪肉是否掺杂,检测结果肉眼可见,灵敏度为10 pg/μL。
Six primers speci fic to the mitochondrial cytochrome b gene of swine were designed for use in loop-mediated isothermal amplification (LAMP). A simplified procedure was used for mitochondrial DNA extraction, and the reaction system was optimized. The specificity was tested with mitochondrial DNA samples from beef, mutton, chicken, duck meat, horse meat, and donkey meat as the negative controls, and the limit of detection (LOD) was determined with gradient dilution of adulterated meat with pork. A visual LAMP method for the detection of pork components in common meat products was established. The results showed that the modified mitochondrial DNA extraction method was simple and repeatable, and the visual LAMP method with 4-(2-pyridylazo)-resorcinol sodium salt as the indicator could accurately detect pork adulteration in common meat products with an LOD of 10 pg/μL.
作者
朱凯
康怀彬
王德国
ZHU Kai;KANG Huaibin;WANG Deguo(College of Food and Bioengineering,Henan University of Science and Technology,Luoyang 471000,China;College of Food and Biological Engineering,Xuchang University,Xuchang 461000,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2019年第12期296-302,共7页
Food Science
基金
“十三五”国家重点研发计划重点专项(2016YFD0500704-4)
2015河南省高等学校科技创新团队支持计划项目(15IRTSTHN016)
2018河南省科技计划项目(182102110285)
关键词
环介导等温扩增
猪肉
检测
线粒体DNA提取
可视化
loop-mediated isothermal ampli fication
pork
detection
mitochondrial DNA extraction
visualization
