橙汁饮料中橙与橘成分双重实时荧光定量PCR检测技术

Dual real-time fluorescence quantitative PCR method for the detection of orange and citrus components in orange juices

目的:建立双重实时荧光定量PCR检测橙汁饮料中橙与橘成分的方法。方法:通过设计和筛选橙、橘成分的引物和探针,检测水果寻找橙和橘的荧光扩增规律,利用橙和橘在FAM/VIC两个荧光通道上的比值差异来判定橙、橘成分。采用模拟橙汁掺杂柑橘汁试验验证方法检测的最低掺杂比例,对市售橙汁饮料进行检验来验证方法可行性。结果:纯橙水果在FAM和VIC两通道均有荧光扩增曲线,FAM/VIC通道荧光比值在0.5±0.2的范围内,而纯橘水果仅在FAM通道有很强的荧光扩增曲线。模拟掺杂试验中,可以检测出橙汁中掺有10%及以上的柑橘汁。结论:该方法能够检测橙汁饮料中的橙和橘成分,可用于市售橙汁饮料的鉴伪。

Objective:To establish a dual real-time PCR method for the detection of orange and citrus components in orange juices.Methods:Through the design and screening of primers and probes for orange and citrus components.Through the detection of fruits,found the rule of fluorescence amplification of orange and citrus.Using the ratio difference of orange and citrus in FAM/VIC two fluorescence channels to determine the composition of orange and citrus.The lowest doping ratio detected by the method was verified by the experiment of orange juice adulteration,tested the commercial orange juice and verify the feasibility of the method.Results:There were fluorescence amplification curves in both FAM and Vic channels in pure orange fruits,and the fluorescence ratio of FAM/VIC channel was between 0.5±0.2,while only FAM channel had strong fluorescence amplification curve in pure citrus fruits.In the simulation experiment,it could be detected that there are more than 10%citrus juice in orange juice.Conclusion:The results showed that this method can be used to detect the orange and citrus components in commercial orange juices,and can be used to identify the fake orange juices.