摘要
目的建立重组酶介导等温扩增技术快速检测猪肉中猪源性成分的方法。方法应用重组酶聚合酶等温扩增技术(recombinase polymerase amplification,RPA),根据猪源性线粒体细胞色素b(Cytb)基因序列设计2对RPA引物,筛选了1对可用于扩增的引物,建立基于重组酶介导的等温扩增技术的检测方法,并对该方法进行扩增温度、特异性和灵敏性验证。结果 30℃等温扩增条件下,RPA引物的特异性良好,灵敏性可达0.1ng/μL。结论本研究成功建立了猪源性肉制品的等温扩增方法,该方法快速简便,具有较高的特异性和灵敏性,适用于市售生鲜肉中猪源性成分的快速鉴定。
Objective To establish a method for rapid detection of porcine ingredient in pork by recombinase polymerase amplification assay （RPA）. Methods According to the Cytb gene sequence of porcine, 2 pairs of RPA primers were designed and one of them was specific to amplify. A novel event-specific detection method for pork based on RPA was established. The specificity and sensitivity of this method were respectively performed for validation. Results Primers for RPA had good specificity under the condition of 30 ℃. This method had a high sensitivity of 0.1 ng/gL. Conclusion The RPA method for detection of porcine ingredient in meat is successfully established. This method is rapid and efficient, and has good specificity and sensitivity, which is suitable for the rapid identification of porcine ingredient in meat from the market.
作者
郭燕华
陈遂
王德莲
聂炎炎
吴环
杨静
曾国权
陈景亮
郭新东
GUO Yan-Hua CHEN Sui WANG De-Lian NIE Yan-Yan WU Huan YANG Jing ZENG Guo-Quan CHEN Jing-Liang GUO Xin-Dong(Guangzhou Quality Supervision and Testing Institute, Guangzhou 511447, China)
出处
《食品安全质量检测学报》
CAS
2017年第6期2012-2016,共5页
Journal of Food Safety and Quality
基金
广东省科自然科学基金项目(2016A030310102)
广东省质量技术监督局科技项目(2016CZ11)~~
关键词
等温扩增
快速检测
猪肉
猪源性成分
isothermal amplification
rapid detection
pork
porcine ingredient
