多重荧光定量聚合酶链式反应法检测水牛乳中黄牛源性成分  

Detection of Bovine Derived Components in Buffalo Milk Using Multiplex Fluorescence Quantitative Polymerase Chain Reaction Method

作　　者：梁媛媛 赵娇 崔生熠 刘鹏鹏 LIANG Yuanyuan;ZHAO Jiao;CUI Shengyi;LIU Pengpeng(Zhejiang Fangyuan Test Group Co.,Ltd.,Hangzhou 310018,China;Key Laboratory of Biosafety Detection for Zhejiang Market Regulation,Hangzhou 310018,China;College of Food Science and Technology,Zhejiang Pharmaceutical University,Ningbo 315500,China)

机构地区：[1]浙江方圆检测集团股份有限公司,浙江杭州310018 [2]浙江省市场监管生物安全重点实验室,浙江杭州310018 [3]浙江药科职业大学食品学院,浙江宁波315500

出　　处：《现代食品》2025年第1期150-154,159,共6页Modern Food

基　　金：浙江方圆检测集团股份有限公司自立项目(ZL202321)。

摘　　要：目的:建立多重荧光定量聚合酶链式反应(Polymerase Chain Reaction,PCR)法同时检测水牛乳中水牛、黄牛这2种牛源性成分。方法:设计基于cytb基因的特异性引物和探针,通过优化反应体系的退火温度和循环参数,建立一种能在同一反应体系内快速检测水牛和黄牛源性成分的多重荧光定量PCR方法。同时,对该方法进行特异性、灵敏度、重复性等方法学验证。结果:该方法特异性强,能特异性检出水牛、黄牛源性成分;该方法灵敏度可达核酸浓度1.0×10^(-4) ng·μL^(-1),且重复性较好。结论:该方法操作简单,具有通量高、特异性强、灵敏度高和重复性好等优点,适用于水牛乳样品中水牛、黄牛源性成分的掺假鉴别和快速检测。Objective:To establish a multiplex fluorescent quantitative polymerase chain reaction(PCR)method for simultaneous detection of two bovine derived components,buffalo and cattle,in buffalo milk.Method:Design specific primers and probes based on the cytb gene and optimize the annealing temperature and cycling parameters in order to establish a multiplex fluorescence quantitative PCR method for rapid detection of buffalo and cattle derived components in a single reaction.Meanwhile methodological validation such as specificity,sensitivity and repeatability was performed.Result:This method has strong specificity and can specifically detect components derived from buffalo and cattle.Its sensitivity can reach a nucleic acid concentration of 1.0×10^(-4) ng·μL^(-1),and the repeatability is good.Conclusion:This method has the advantages of simplicity of operation,high flux,strong specificity,and high sensitivity,and is suitable for the identification and rapid detection of buffalo and cattle in adulteration of buffalo milk.

关 键 词：多重荧光定量聚合酶链式反应 水牛乳 牛源性成分 快速检测 

分 类 号：O657.34[理学—分析化学]