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1 5 2 Vol. 5 No Journal of Food Safety and Quality Feb., 2014 PCR 郭凤柳, 熊蕊, 刘晓慧 *, 赵同欣, 王娜, 颜红 (, ) 摘要 : 目的 5 PCR, 4 PCR, 方法 DNA, PCR,, 结果,, 56, 75%; 15, 6.7%; 5, 20%; 2, 1,, 结论 PCR DNA, pg fg, 关键词 : PCR ; ; Application of PCR technique for the detection of meat source components GUO Feng-Liu, XIONG Rui, LIU Xiao-Hui *, ZHAO Tong-Xin, WANG Na, YAN Hong (Baoding Entry-Exit Inspection and Quarantine Bureau, Baoding , China) ABSTRACT: Objective To establish an ordinary PCR method to detect 5 kinds of animal origin ingredients, a real time fluorescent PCR method to detect 4 kinds of animal origin ingredients, and serve the food safety. Methods Genomic DNA were extracted from the meat, PCR detection primers and fluorescent probes were synthesized, and the reaction conditions and reaction system were optimized to establish the detection method of animal origin, so as to analyze the meat adulteration in market. Results The measuring method for several kinds animal origin ingredients was established. According to the established method, the adulteration rate for lamb products, donkey products, beef products, and rabbit meat products collected from Baoding area were 75%, 6.7%, 20%, 50%, respectively. The results showed that the adulteration problem existed in Baoding area market, especially in the free market and barbecue samples. The adulteration was serious damage to consumers rights. Conclusion The established PCR method can obtain DNA quickly, its detection sensitivity can achieve pg or fg level, and is suitable for detecting a large number of meat products. KEY WORDS: PCR detection method; detection; adulteration, GB (GB ),, 基金项目 : (HE2011K019) Fund: Supported by Autonomy Projects of Baoding Entry-Exit Inspection and Quarantine (HE2011K019) * 通讯作者 :,, drlxh@163.com *Corresponding author: LIU Xiao-Hui, PhD, Baoding Entry-Exit Inspection and Quarantine Bureau, Baoding , China. drlxh@163.com

2 542 5 [1],,, [2,3],,,,,,, PCR, ; PCR, 1 材料与方法 1.1 试剂及仪器 : K RNA dntps 10 PCR MgCl 2 Taq DNA 2000 bp ladder DNA marker / / DNA DNA Sau3AⅠ HphⅠ MnlⅠ AluⅠ, : ABI PCR PCR ( ( ) ) QUANTUM-ST4 ( ) ( ( ) ) Sigma ( ) 1.2 检测方法 普通 PCR 法 DNA PCR, DNA,, DNA,,, : DNA 10 1 ~10 6,, DNA,,,, 实时荧光 PCR 法 : PCR ( Real Time PCR Bovine and Ovine DNA Detection Kit Real Time PCR Porcine DNA Detection Kit ) BioXL (Duck) (PCR- ),,, 25 μl 10 PCR buffer 9 μl Mg μl dntps 2 μl (10 μmol/l)1 μl (10 μmol/l)1 μl DNA(1~100 ng/μl)1 μl 9.5 μl : s, 95 5 s s 检测方法的应用, DNA 56 ; 10, 5 ; 2 结果与分析,,

3 2, : PCR 543 DNA pg/μl, fg/μl, PCR DNA, 56, 38,, 18, 11, 7 1 1, 14, 75%, 10, ; 5, 1 ;, 5, 1, 3,,, %,, 3 讨论,,,,,,,, Table 1 表 1 市售羊肉串 ( 卷 ) 检测结果 Commercial lamb skewers (volume) test tesults % Fig. 1 Adulteration percentage column chart

4 544 5 Table 2 表 2 小作坊 小吃摊 正规超市及饭店肉制品掺假数比较 Comparison of relatively adulterated meat among small workshops, food stalls, supermarkets and hotels Fig. 2 Comparison of relatively adulterated meat among small workshops, food stalls, supermarkets and hotels,,,,,, DNA,, DNA [4],, PCR, [5] [6] DNA DNA, DNA,, DNA [7,8] ,,, PCR DNA Ali RT-PCR, TaqMan probe,, 0.01% [9] PCR, PCR, PCR,,, 2011 PCR, DNA 98 fg/μl DNA 67 fg/μl, [10], DNA, PCR DNA, pg fg,,,,, 参考文献 [1]. [D]., 2005.

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