肉串制品中豬牛羊源性成分多重熒光PCR檢測方法研究

摘要 ［目的］建立畜類肉串產品中常見的黃牛、綿羊和豬源性成分的多重熒光PCR檢測方法，實現肉串制品中動物源性成分的快速鑒別。［方法］比較磁珠法和柱膜法對肉糜組織中DNA的提取效果，優化實時熒光PCR多重檢測方法，比較58、60 ℃ 2個退火溫度下黃牛、綿羊、豬源性成分擴增效果。［結果］試驗采用的2種DNA提取方法效果相似，DNA溶液的A260/A280值均能滿足PCR檢測要求;58 ℃為最優退火溫度，此時黃牛、綿羊、豬源性成分均能有效擴增，且最低檢出的DNA濃度均為10-3 ng/μL。方法中用到的引物和探針特異性較好，均未產生非特異性擴增。［結論］試驗建立的黃牛、綿羊和豬源性成分的多重PCR檢測方法能為肉制品中動物源性成分的快速定性分析提供方法參考。

關鍵詞 黃牛;綿羊;豬;源性成分;實時熒光多重PCR

中圖分類號 TS251.7 文獻標識碼 A

文章編號 0517-6611（2024）18-0184-04

doi：10.3969/j.issn.0517-6611.2024.18.039

開放科學（資源服務）標識碼（OSID）：

Study on Detection Method for Pig，Cow and Sheep Derived Components by Multiple Fluorescence PCR in Meat Skewer Products

YAO Yan-ling，ZHOU Yu-dong，WANG Wen-yu et al

（Jiaxing Testing Institute for Food，Drug and Product Quality Control，Jiaxing，Zhejiang 314050）

Abstract ［Objective］To establish a multiple fluorescence PCR detection method for common cattle，sheep and pig derived components in animal meat skewer products，and to achieve rapid identification of animal derived components in meat skewer products.［Method］Compared the extraction effect of DNA from meat mince tissue using magnetic bead method and column membrane method，optimized the real-time fluorescence PCR multiple detection method，and compared the amplification effect of cattle，sheep，and pig derived components at two annealing temperatures of 58 and 60 ℃.［Result］The two DNA extraction methods used in the experiment had similar effects，and the A260/A280 values of the DNA solution could meet the requirements of PCR detection.58 ℃ was the optimal annealing temperature，at which point the cattle，sheep and pig derived DNA could be effectively amplified，and the lowest detected DNA concentration was 10-3 ng/μL.The primers and probes used in the method had good specificity and had not produced non-specific amplification.［Conclusion］The multiplex PCR detection method established by the application scope experiment for cattle，sheep and pig origin can provide a method reference for rapid qualitative analysis of animal origin components in meat products.

Key words Cattle;Sheep;Pig;Derived component;Real-time fluorescence multiplex PCR

基金項目 浙江省市場監督管理局雛鷹計劃培育項目（CY2023322）。

作者簡介 姚艷玲（1980—），女，浙江嘉興人，高級工程師，碩士，從事食品微生物及分子生物檢測研究。

收稿日期 2023-10-28

燒烤食品近年來備受年輕人喜愛，尤其以豬、牛、羊等畜肉類肉串最為常見，因其含有多種維生素、氨基酸，且燒烤口味鮮美，深受年輕人青睞，也豐富了人們的餐桌文化。隨著市場上豬肉、牛肉和羊肉價格差距的拉大，某些商販在肉制品加工過程中摻假售假，通過牛羊油浸泡，或者牛羊類香精香料及調味料添加等方式處理后，冒充牛、羊肉進行銷售，損害了消費者利益，也給食品安全帶來了風險。……