重组酶介导等温扩增技术快速检测牛肉及 牛肉制品中的牛源性成分

Determination of bovine ingredient in beef and its derivates with recombinase polymerase mediated isothermal amplification

中文关键词:  等温扩增  快速检测  牛肉  牛源性成分

英文关键词:isothermal amplification  rapid detection  beef  bovine ingredient

中文摘要:

      目的 建立重组酶介导等温扩增技术快速检测牛肉及牛肉制品中牛源性成分的方法。方法 应用重组酶聚合酶等温扩增技术(recombinase polymerase amplification, RPA), 根据牛源性线粒体细胞色素b(Cytb)基因序列设计筛选了一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 40 ℃等温扩增条件下, 所设计的引物的特异性为100%, 该检测方法的灵敏性可达0.1 ng/μL。结论 本研究成功建立了牛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, 适用于市售生鲜肉制品及加工肉制品中牛源性成分的鉴定。

英文摘要:

      Objective To establish a method for rapid detection of bovine ingredient in beef and its derivates by recombinase polymerase amplification assay (RPA). Methods According to the Cytb gene sequence of bovine, a pair of RPA primers was designed and a novel event-specific detection method for beef and its derivates 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 40 ℃. This method had a high sensitivity of 0.1 ng/?L. Conclusion The RPA method for detection of bovine in meat and meat products is successfully established. This method is rapid, efficient and suitable for the identification of meat and its derivates from the market.