不同核酸提取方法用于多重荧光PCR法检测3种混合熟肉的比较分析

Comparison and analysis of different nucleic acid extraction methods for detection of 3 kinds of mixed cooked meat by multiple fluorescence PCR

目的探讨多重荧光定量PCR法检测不同核酸提取方法以及蒸、煮、烤烹饪方式制作的混合熟肉制品的差异。方法用3种不同的提取方法:抽提法、离心柱法及磁珠法,提取经过蒸、煮、烤烹饪方式制作的牛、鸡、猪、鸭混合样品的DNA,比较不同方法提取DNA的质量及多重荧光PCR检测提取的DNA的效果。结果 3种方法提取DNA的浓度及纯度无明显差别,多重实时荧光PCR检测磁珠法提取的混合样品DNA的Ct值最小,扩增效果最佳。结论本研究中磁珠法提取熟肉制品DNA的检测效果更为理想。

Objective To investigate the difference of multiple fluorescence quantitative PCR detection results of different nucleic acid extraction methods and steamed, cooked and roasted cooked meat products. Methods Three different extraction methods including extraction, centrifugal column and magnetic beads were used to extract DNA from the mixed samples of cattle, chicken, pig and duck prepared by steaming, boiling and baking. The quality of DNA extracted by different methods and the effect of DNA extraction for multiplex PCR detection were compared. Results There was no significant difference in the concentration and purity of DNA extracted by the 3 methods. The Ct value of DNA extracted by magnetic bead method was the smallest and the amplification effect was the best. Conclusion In this study, the detection effect of DNA extracted from cooked meat products by magnetic beads method is more ideal.