基于PCR和LFS技术的掺假豌豆成分快速检测

Rapid detection of adulterated pea components based on PCR and LFS technology

文章运用聚合酶链式反应(polymerase chain reaction,PCR)和侧向流试纸条(lateral flow strip,LFS)相结合方法,以豌豆特异性基因为参照对绿豆食品中掺入豌豆成分进行定性分析。实验对退火温度、修饰引物浓度和镁离子浓度进行优化,并在最优PCR条件下进行特异性和灵敏度检测。结果表明,在退火温度为59℃,修饰引物加入量为0.20μmol/L,镁离子加入量为1.75 mmol/L条件下,PCR扩增效果最好,掺假检测限为0.5%,最低可检测豌豆DNA量为0.12 ng。该方法特异性高、操作简单快速,依靠视觉观察就能得出定性结果。

In this study,polymerase chain reaction(PCR)and lateral flow strip(LFS)were used in the qualitative analysis of pea ingredients in mung bean food with reference to pea specific genes.The annealing temperature,modified primer concentration and magnesium ion concentration were optimized,and the specificity and sensitivity of the samples were detected under the optimal PCR condition.The results showed that under the conditions of annealing temperature of 59°C,modified primer concentration of 0.20μmol/L and magnesium ion concentration of 1.75 mmol/L,the PCR amplification effect was the best,the adulteration limit of detection was 0.5%,and the minimum detectable amount of pea DNA was 0.12 ng.The method has high specificity,simple and fast operation,and qualitative results can be obtained by naked eye observation.