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目的:研究SYBR Green Ⅰ实时荧光PCR检测食品中转基因成分(35S基因和NOS基因)方法的灵敏度、特异性和线性关系。方法:使用SYBR Green Ⅰ实时荧光PCR方法检测不同转基因含量的标准品,并对转基因阳性和阴性样品分别进行扩增,分析扩增曲线和Ct值。结果:SYBR Green Ⅰ实时荧光PCR检测食品中转基因成分方法检出限为0.05%,转基因大豆标准品和阳性样品中检出35S基因和NOS基因成分,阴性样品中未检出35S基因和NOS基因成分。结论:SYBR Green Ⅰ实时荧光PCR检测35S和NOS转基因成分方法灵敏度高,特异性较好,标准曲线线性良好。
OBJECTIVE: To study the sensitivity, specificity and linearity of SYBR Green Ⅰ real-time fluorescence PCR for detecting the content of 35S gene and NOS gene in food. Methods: The SYBR Green Ⅰ real-time fluorescent PCR method was used to detect the standard content of different gene content. The positive and negative transgenic samples were respectively amplified, and the amplification curve and Ct value were analyzed. Results: The detection limit of SYBR Green Ⅰ real-time PCR was 0.05%. 35S gene and NOS gene were detected in transgenic soybean standard samples and positive samples, but 35S gene and NOS gene were not detected in negative samples . Conclusion: SYBR Green Ⅰ real-time fluorescence PCR detection of 35S and NOS transgenic methods has high sensitivity, good specificity and good linearity of standard curve.