微滴式数字PCR定量检测欧洲甜樱桃中掺假苹果成分的研究

目的:基于微滴式数字PCR技术（Droplet Digital PCR，ddPCR）建立欧洲甜樱桃中掺假苹果成分定量检测的方法。方法:通过筛选欧洲甜樱桃、苹果成分特异性引物探针，建立ddPCR定量检测方法，构建质量和DNA含量的拟合曲线及DNA含量与拷贝数之间的拟合曲线，以DNA含量作为中间值，获得质量和扩增拷贝数的计算公式。其中线性拟合曲线R2均在0.99以上，且建立了相应的掺假模型以验证方法的准确性。结果:欧洲甜樱桃和苹果的质量和扩增DNA拷贝数的计算公式:M_樱=0.0833C?3.8420、M_苹=0.4084C?1.5747。在欧洲甜樱桃与苹果的掺假模型中相对误差最大为?19.17%，符合统计学要求。结论:建立欧洲甜樱桃制品快速检测方法为打击欧洲甜樱桃掺杂使假提供了强有力手段，保障了消费者的权益，对促进小浆果及其制品行业健康发展具有重要意义。

Quantitative Detection of Adulterated Apple Components in European Sweet Cherries by Droplet Digital PCR

Objective: To establish a method for quantitative detection of adulterated apple components in cherry based on droplet digital PCR (ddPCR). Methods: By screening the component specific primer probes of cherry and apple, the quantitative detection method of ddPCR was established. The fitting curve between quality and DNA content and the fitting curve between DNA content and copy number were constructed. Taking DNA content as the intermediate value, the calculation formulas of quality and amplified copy number were obtained. The linear fitting curves R2 were all above 0.99, and the corresponding adulteration model was established to verify the accuracy of the method. Results: The formula for calculating the mass and amplified DNA copy number of cherry and apple: M_cherry=0.0833C?3.8420, M_apple=0.4084C?1.5747. In the adulteration model of cherries and apples, the maximum relative error was ?19.17%, which met the statistical requirements. Conclusion: The establishment of a rapid detection method for European sweet cherry products provides a powerful means to combat the adulteration of European sweet cherries, protects the rights and interests of consumers, and is of great significance for promoting the healthy development of the small berry and its product industry.