基于重组酶等温扩增技术快速检测生鲜肉中猪源性成分

Determination of porcine ingredient in pork with recombinase polymerase mediated isothermal amplification

中文关键词:  等温扩增  快速检测  猪肉  猪源性成分

英文关键词:isothermal amplification  rapid detection  pork  porcine ingredient

中文摘要:

      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0.1 ng/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。

英文摘要:

      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/?L. 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.