摘要
探讨采用实时荧光定量PCR方法检测食品中的花生和芹菜成分。实验结果表明;采用花生引物2和探针2进行PCR扩增时,27种样品只有花生产生荧光信号,其余样品均不产生荧光信号;采用芹菜的引物和探针进行扩增时,18种样品只有芹菜和西芹产生荧光信号,其余样品均不产生荧光信号。灵敏度实验结果表明:花生能检测到10-5稀释度的3.6pgDNA的量,定量关系式为:y=-3.092261x+21.216097,R2=0.993068。芹菜能检测到10-5稀释度的4.0pgDNA的量,定量关系式为:y=-4.0957441x+23.412189,R2=0.997039。结果表明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。
The USA and the EU have legislated to require labeling of foods containing allergic ingredients. There are eight kinds of main allergic ingredients, including peanut and celery. A real-time fluorescent PCR assay was presented for the quantitative detection of peanut and celery ingredients in foods in the present study. When using the second pair of primer and probe （a total of two pairs were designed） of peanut for PCR amplification, among amplified DNA extracted from 27 samples only peanut produced fluorescent signal. Likewise, when using a pair of prime and probe designed using celery DNA for PCR amplification, only Chinese celery and western celery produced fluorescent signal in 18 samples after real-time PCR amplification. The sensitivity test indicated that 3.6 pg of peanut DNA could be detected, with a quantitative formula： y = -3.092261x ＋ 21.216097 （R2= 0.993068）, where y was Ct, and x was natura/logarithm of the DNA concentration （the same below）; and 4.0 pg of celery DNA could be detected, with a quantitative formula： y = - 4.0957441x ＋ 23.412189 （R^2 = 0.997039）. The results demonstrate that these pairs of prime and probe have high sensitivity and amplification efficiency and therefore can meet the requirement of trace detection of peanut and celery ingredients in foods.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2009年第14期261-263,共3页
Food Science
