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根据鸭mtDNA COX基因上的保守序列设计特异性引物和TaqMan探针,建立实时荧光定量PCR用于检测畜禽肉制品中的鸭源性成分。结果表明,建立的方法特异性强,与鹅、鸡、羊、牛等15种动物DNA无非特异性扩增;灵敏度高,可检测1.0pg/μL鸭源DNA的存在;重复性好,同DNA浓度所测得Ct值的变异系数均小于3%;应用该法对模拟混合样品进行检测,结果与预期相符,且常见肉类DNA的存在并不影响该法对鸭源性成分检测的灵敏度。说明本试验建立的鸭源性成分实时荧光定量PCR法特异、敏感、稳定,可快速准确检测畜禽肉制品中含有的鸭源性成分。
Based on the conserved sequence of duck mtDNA COX gene, specific primers and TaqMan probes were designed and used to detect duck-derived components in poultry and meat products by real-time fluorescence quantitative PCR. The results showed that the established method was highly specific and non-specific amplification with DNA from 15 species of goose, chickens, sheep and cattle. The sensitivity was high, which could detect 1.0pg / The coefficient of variation (Ct) was less than 3%. The method was applied to simulate mixed samples and the results were in accordance with the expectation. The presence of common meat DNA did not affect the sensitivity of this method to the detection of duck origin components. This study established that the duck-derived components of real-time fluorescence quantitative PCR method is specific, sensitive and stable, and can quickly and accurately detect the duck-derived components contained in livestock and poultry meat products.