乳业科学与技术 ›› 2023, Vol. 46 ›› Issue (6): 20-24.DOI: 10.7506/rykxyjs1671-5187-20231224-062

• 分析检测 • 上一篇    下一篇

基于实时荧光定量技术建立定量检测乳粉中羊源性成分的方法

陈 晨,张 瑞,王玉欣,张同贺,张子仑,贾文珅,周 巍   

  1. 1.河北省食品检验研究院,河北省食品安全重点实验室,国家市场监管重点实验室(特殊食品监管技术),特殊食品安全与健康河北省工程研究中心,河北 石家庄 050227;2.北京市农林科学院,北京 100050
  • 发布日期:2024-03-27
  • 基金资助:
    河北省市场监督管理局科研计划项目(2022ZD11)

Establishment of a Fluorescence-Based Quantitative Real-Time PCR Method for Detection for Caprine-Derived Ingredients in Milk Powder

CHEN Chen, ZHANG Rui, WANG Yuxin, ZHANG Tonghe, ZHANG Zilun, JIA Wenshen, ZHOU Wei   

  1. 1. Hebei Food Safety Key Laboratory, Key Laboratory of Special Food Supervision Technology for State Market Regulation, Hebei Engineering Research Center for Special Food Safety and Health, Hebei Food Inspection and Research Institute, Shijiazhuang 050227, China; 2. Beijing Academy of Agricultural and Forestry Sciences, Beijing 100050, China
  • Online:2024-03-27

摘要: 基于实时荧光定量聚合酶链式反应建立了乳粉中羊源性成分相对定量检测方法。对羊引物进行特异性、灵敏度和稳定性实验。通过模拟不同质量浓度羊乳粉与马乳粉混合样本,根据其ΔCt值的函数关系进行线性拟合进而绘制标准曲线,建立乳粉中羊源性成分相对定量检测。结果显示,该方法最低检测限为0.000 1 ng/μL、标准曲线方程为y=0.720 9x+5.651 9(R2=0.987 5)、回收率为96.22%~112.00%、组间变异系数≤0.79%、组内变异系数≤1.77%。

关键词: 羊乳粉;马乳粉;实时荧光定量聚合酶链式反应;掺假检测

Abstract: This study established a fluorescence-based quantitative real-time polymerase chain reaction method for the detection of caprine-derived ingredients in milk powder. The specificity, sensitivity, and stability of the caprine-specific primers were evaluated. By linear fitting of the difference in cycle threshold (ΔCt) as a function of the mixing proportion between goat and horse milk powder, a calibration curve for the relative quantitation of caprine-derived ingredients in milk powder was established as follows: y = 0.720 9x + 5.651 9 (R2 = 0.987 5), and the minimum detection limit of this method was 0.000 1 ng/μL. The recoveries of the proposed method were 96.22%-112.00%, and the inter- and intra-group coefficient of variation was ≤ 0.79% and ≤ 1.77%, respectively.

Key words: goat milk powder; horse milk powder; real-time fluorescence quantitative polymerase chain reaction; adulteration detection

中图分类号: