实时荧光定量PCR鉴定食品中绿豆源性成分

Identification of mung bean components in food by real-time fluorescence quantitative PCR

通过设计和筛选引物、探针组合,优化实时荧光定量PCR(qPCR)反应体系,建立了绿豆源性成分qPCR鉴定方法,并对其准确性进行了验证。结果表明,筛选出的引物、探针组合特异性良好,通过对其反应终浓度和退火温度的优化,获得了最优的qPCR反应条件,扩增效率可达100.3%。确定了绿豆源性成分的检出限为0.05%,定量限为0.1%。在实际应用中,模拟样品、市售样品的定量值标准差和相对标准偏差均小于25%。由此可知,建立的实时荧光定量PCR方法能够特异性鉴定食品中的绿豆源性成分,并对其进行定量,准确度较高。

By designing and screening primer and probe combinations,the real-time fluorescence quantitative PCR(qPCR)reaction system was optimized,and the qPCR method was established to identify the source components of mung bean,and its accuracy was verified.The results showed that the combination of primers and probes screened had good specificity.By optimizing the final concentration and annealing temperature,the optimal qPCR reaction conditions were obtained,and the amplification efficiency could reach 100.3%.The detection limit and quantification limit of mung bean were 0.05%and 0.1%,respectively.In practice,the quantitative deviation and relative standard deviation of simulated samples and commercial samples were less than 25%.The established real-time fluorescence quantitative PCR method could specifically identify and quantify mung bean components in food,with high accuracy.