食品和饲料中鹅源性成分微滴式数字PCR检测与定量分析

Detection and Quantification of Goose-derived Materials in Food and Feedstuffs by Droplet Digital PCR

摘要:

本研究利用微滴式数字PCR（droplet digital polymerase chain reaction，ddPCR）技术对食品和饲料中鹅源性成分进行检测与精确定量。选取鹅的单拷贝核基因作为靶基因，建立了鹅源性成分靶基因拷贝数与样品质量之间的线性关系；选取21种常见动物物种及鹅的9个品系对所设计的引物及探针的种间特异性和种内保守性进行了验证，并对该方法的检测下限（limit of detection，LOD）和定量检测下限（limit of quantification，LOQ）进行了验证，利用已知鹅源性成分含量的样品和市售商品对该方法的准确性和适用性分别进行了验证。结果表明，所设计的引物及探针具有良好的种间特异性和种内保守性，该定量检测方法的检测下限和定量检测下限分别为1 copy/μL和5 copies/μL，且该方法具有良好的准确性和适用性，可用于对食品和饲料中鹅源性成分进行检测和精确定量。

Abstract:

In this study, a highly precise, quantitative method based on the droplet digital polymerase chain reaction (ddPCR) technique was developed to identify and quantify the goose content in food and feedstuffs. The single-copy Anser cygnoides domesticus genomic scaffold gene was selected as the target gene, a formula for calculating raw meat weight based on target DNA copy number was established. Exclusive specificity was verified using samples from 21 different animal species, and inclusive specificity of goose was tested using nine different breeds. The limit of detection (LOD) and the limit of quantification (LOQ) of the method were 1 and 5 copies/μL, respectively. The accuracy and applicability of the method were verified using mixed powder samples with known proportions and commercial products.The results confirmed that the developed ddPCR method is highly precise for identifying and quantifying the goose content in food and feedstuffs, indicating the potential applicability in future routine analyses.