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基于针式扎取法与苯酚-氯仿抽提法的牦牛肉及常见畜禽肉DNA提取效率与质量对比
普巴扎西1，扎西穷达1，刘亚洲1，米玛1，侯智轩2，张朵朵2，尼珍1*，刘永峰2*
(1.西藏自治区食品药品检验研究院，西藏功能食品与食品质量安全重点实验室，西藏 拉萨 850000；2.陕西师范大学 食品工程与营养科学学院，陕西 西安 710119)
Comparative Study of DNA Extraction Efficiency and Quality between Needle Pricking Method and Phenol-Chloroform Extraction Method in Yak Meat and Common Livestock and Poultry Meat
Puba Zhaxi1， Zhaxi Qiongda1， LIU Yazhou1， Mi Ma1， HOU Zhixuan2， ZHANG Duoduo2，Ni Zhen1*， LIU Yongfeng2*
(1. Xizang Institute for Food and Drug Control， Xizang Key Laboratory of Functional Food and Food Quality and Safety， Lhasa 850000， Xizang， China； 2. College of Food Engineering and Nutritional Science， Shaanxi Normal University， Xi′an 710119， Shaanxi， China)
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投稿时间：2025-07-21
中文摘要: 为满足现场快速掺假检测的需求，该研究分别以牦牛肉、鸡肉、猪肉、鸭肉和牛肉的生鲜肉和肉制品为样本，以针式扎取法提取肉品DNA，与传统的苯酚-氯仿抽提法进行比较，探究不同钨针数量对所提DNA质量的影响，并利用实时荧光定量聚合酶链式反应（quantitative real-time polymerase chain reaction，qPCR）建立的灵敏度标准曲线计算针式扎取法提取的DNA浓度。结果 表明，钨针扎取法提取生鲜肉DNA的A260/A280为1.10～1.35，肉制品DNA的A260/A280为1.40～1.60，其浓度为3～1 300 pg/μL，提取的DNA可用于后续qPCR检测。且该方法可在2 min内提取出适用于后续检测的DNA，极大缩短提取时间，且成本相对较低。
Abstract:To meet the demand for rapid on-site adulteration detection， this study used fresh yak meat，chicken， pork， duck， and beef， as well as their meat products， as samples. DNA was extracted from these samples using the needle pricking method and compared with the traditional phenol-chloroform extraction method. The impact of different numbers of tungsten needles on the quality of extracted DNA was investigated，and the concentration of DNA extracted by the needle puncture method was calculated using the sensitivity standard curve established by quantitative real-time polymerase chain reaction （qPCR）. The results indicated that the A260/A280 value of DNA extracted from fresh meat by the tungsten needle pricking method was 1.10-1.35， and that of meat products was 1.40-1.60. The concentration ranged from 3 to 1 300 pg/μL. The extracted DNA could be used for subsequent qPCR detection. Moreover， this method could extract DNA suitable for subsequent detection within two minutes， significantly reducing the DNA extraction time， with relatively low cost.
文章编号：202601018 中图分类号： 文献标志码：
基金项目:2024 年度西藏自治区科技厅重点研发计划项目（XZ202402ZY0006）
| 作者 | 单位 |
| 普巴扎西 | 1.西藏自治区食品药品检验研究院,西藏功能食品与食品质量安全重点实验室,西藏 拉萨 850000； |
| 扎西穷达 | 1.西藏自治区食品药品检验研究院,西藏功能食品与食品质量安全重点实验室,西藏 拉萨 850000； |
| 刘亚洲 | 1.西藏自治区食品药品检验研究院,西藏功能食品与食品质量安全重点实验室,西藏 拉萨 850000； |
| 米玛 | 1.西藏自治区食品药品检验研究院,西藏功能食品与食品质量安全重点实验室,西藏 拉萨 850000； |
| 侯智轩 | 2.陕西师范大学 食品工程与营养科学学院,陕西 西安 710119 |
| 张朵朵 | 2.陕西师范大学 食品工程与营养科学学院,陕西 西安 710119 |
| 尼珍 | 1.西藏自治区食品药品检验研究院,西藏功能食品与食品质量安全重点实验室,西藏 拉萨 850000； |
| 刘永峰 | 2.陕西师范大学 食品工程与营养科学学院,陕西 西安 710119 |
| Author Name | Affiliation |
| Puba Zhaxi | 1. Xizang Institute for Food and Drug Control, Xizang Key Laboratory of Functional Food and Food Quality and Safety, Lhasa 850000, Xizang, China； |
| Zhaxi Qiongda | 1. Xizang Institute for Food and Drug Control, Xizang Key Laboratory of Functional Food and Food Quality and Safety, Lhasa 850000, Xizang, China； |
| LIU Yazhou | 1. Xizang Institute for Food and Drug Control, Xizang Key Laboratory of Functional Food and Food Quality and Safety, Lhasa 850000, Xizang, China； |
| Mi Ma | 1. Xizang Institute for Food and Drug Control, Xizang Key Laboratory of Functional Food and Food Quality and Safety, Lhasa 850000, Xizang, China； |
| HOU Zhixuan | 2. College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi′an 710123, Shaanxi, China |
| ZHANG Duoduo | 2. College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi′an 710124, Shaanxi, China |
| Ni Zhen | 1. Xizang Institute for Food and Drug Control, Xizang Key Laboratory of Functional Food and Food Quality and Safety, Lhasa 850000, Xizang, China； |
| LIU Yongfeng | 2. College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi′an 710126, Shaanxi, China |
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