实时荧光PCR法检测食品中芝麻过敏原成分

Real-time Fluorescent PCR Detection of Allergen in Sesame

本实验探讨采用实时荧光定量PCR方法检测食品中的芝麻成分,结果表明:采用芝麻的引物和探针进行扩增时;22种样品经实时PCR扩增,只有白芝麻和黑芝麻产生荧光信号,其余样品均不产生荧光信号。灵敏度实验结果表明能检测到10-5稀释度的4.4×10-12gDNA的量,定量关系式为:y=-3.472018x+18.851009,R2=0.993088。这两对引物和探针具有极高的灵敏度和扩增效率,符合痕量检测要求。

This study aimed to develop a real-time fluorescent PCR assay for specific detection of allergen in sesame. Real- time PCR amplification was conducted to DNA fragments from 22 samples including white sesame and black sesame as well as other different samples using a primer pair and a probe specific for sesame DNA fragment. Both the black sesame and white sesame DNA fragments exhibited fluorescent signal, but others did not. Sensitivity analysis showed that 4.4×10^-12g of sesame DNA with the dilution of 10^-5 could be detected, and the quantitative formula was Ct=-3.4720181 In （initial copy number） ＋ 18.851009, R^2 = 0.993088, indicating that the primer pair and the probe have high sensitivity and amplification efficiency and fit with the requirements of trace detection.