实时荧光定量PCR检测肉制品中牛源性成分及其含量

谭波; 杨春萍; 刘小玲

doi:10.13651/j.cnki.fjnykj.2023.10.002

实时荧光定量PCR检测肉制品中牛源性成分及其含量

谭波^1,2,
杨春萍³,
刘小玲^1, ,

1. 广西大学轻工与食品工程学院, 广西南宁 530004;
2. 贵州省检测技术研究应用中心, 贵州贵阳 550014;
3. 贵州省分析测试研究院, 贵州贵阳 550014

基金项目:

贵州省科技计划项目(黔科合支撑〔2023〕一般354)。

详细信息

作者简介:
谭波,男,1991年生,硕士研究生,工程师,主要从事食品加工与安全研究。

通讯作者:
刘小玲,女,1972年生,博士,教授,主要从事食品生物技术研究(E-mail:1106133912@qq.com)。

中图分类号: TS 251.5
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出版历程
- 收稿日期: 2023-09-19
- 刊出日期: 2023-10-27

Detection of the Bovine-derived Materials and Their Contents in Meat Products by Real-time Fluorescence Quantitative PCR

1. College of Light Industry and Food Engineering, Guangxi University, Nanning, Guangxi 530004, China;
2. Guizhou Research and Application Center of Detection Technology, Guiyang, Guizhou 550014, China;
3. Guizhou Institute of Analysis and Testing, Guiyang, Guizhou 550014, China

摘要

摘要: 建立一种快速、高效、准确且低成本的肉制品中牛源性成分鉴定及量化检测方法。以细胞核单拷贝基因bosPDE为目的基因,LcoR为内参基因分别合成引物及探针,优化反应条件,评价该方法的特异性和灵敏度。采用实时荧光PCR相对定量法,以△Ct值与2^△△Ct值线性模型分别建立回归曲线,通过人工模拟混合牛肉样品对方法准确性评估。结果表明:目的基因bosPDE引物仅对牛肉DNA进行扩增,具有特异性,对牛肉DNA扩增的灵敏度可达0.01 ng·μL^-1。以△Ct值与2^△△Ct值为定量指标的回归曲线分别为y=-3.096 5x+8.479 7(R²= 0.998)、y=0.712 2x+3.050 4(R²=0.988 7)。两个方法检测35%~90%牛肉含量的人工模拟混合牛肉样品的回收率为分别为94.30%~102.52%、98.55%~106.30%。将两种定量方法进行数均处理,回收率的偏差减少。本研究建立的实时荧光定量PCR方法能够准确对肉制品中的牛源性成分进行定性,并可用于牛肉掺伪制品中的牛肉相对定量分析,对鉴别肉制品掺假以及量化牛肉制品成分含量具有重要参考价值。
- 肉制品 /
- 牛源性成分 /
- 实时荧光定量PCR /
- 相对定量
Abstract: In order to establish a rapid, efficient, accurate and low-cost method for the identification and quantitative detection of the bovine-derived materials in meat products, the primers and probes were synthesized with the single-copy gene of cell nucleus, bosPDE as the target gene and LcoR as the internal reference gene, to optimize the reaction conditions and evaluate the specificity and sensitivity of the method. By using the relative quantitative method of real-time fluorescence PCR, the regression curves were respectively established with the linear model of △Ct value and 2^△△Ct value, and the accuracy of the method was evaluated by simulating artificially the mixed beef samples. The results showed that: the bosPDE primer of the target gene only amplified the DNA of beef, which was specific, and the sensitivity of amplification to beef DNA could reach 0.01 ng·μL^-1. The regression curves with △Ct value and 2^△△Ct value as the quantitative indexes were y=-3.096 5x+8.479 7 (R²=0.998) and y=0.712 2x+3.050 4 (R²=0.988 7), respectively. The recovery rates of the artificial simulated mixed beef samples with 35%-90% beef content detected by the two methods were 94.30%-102.52% and 98.55%-106.30%, respectively. The deviation of recovery rate was reduced when the number-average treatment was carried out on the two quantitative methods. The real-time fluorescence quantitative PCR method established in this study could accurately characterize the bovine-derived materials in meat products, and could be used for the relative quantitative analysis of beef in the beef adulterated products. It had important reference value for identifying the adulteration of meat products and quantifying the concentrations of components in beef products.
- Meat products /
- Bovine-derived materials /
- Real-time fluorescence quantitative PCR /
- Relatively quantitative

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