交叉引物等温扩增检测猪肉源性成分

Cross Priming Amplification with Nucleic Acid Test Strip Analysis of Pork in Meat Mixtures

DOI：10.3969/j.issn.1673-1689.2018.11.014

中文关键词: 交叉引物恒温扩增 核酸试纸条 D-loop基因 猪

英文关键词: cross priming amplification,nucleic acid test strip,D-loop gene,pork

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中文摘要:

肉制品掺假现象已成为我国食品行业重点关注的问题。本研究使用交叉引物恒温扩增技术（Cross priming amplification，CPA）和核酸试纸条检测相结合的方法快速检测肉制品中猪肉成分。对猪线粒体的D-loop基因序列设计特异性的引物和探针，通过优化引物浓度及反应条件，确立最佳反应体系。实验结果表明，CPA扩增系统对猪源性成分的检测特异性良好，对单猪源性成分的检出限为1 ng/μL，对混合肉制品中猪源性成分的检出比例为10%（相当于10 ng/μL）。同时核酸试纸条可以快速检测CPA扩增产物，结果与凝胶电泳系统检测保持一致。本研究建立的CPA-核酸试纸条检测方法是一种特异、高效的肉制品中猪肉成分检测方法，可以为肉制品质量控制提供有效的新手段。

英文摘要:

The phenomenon of meat adulteration has become the concerned problems of the food industry in China. In this study，the cross priming amplification（CPA） and nucleic acid test strip detection method were used together to rapid detect the component of pork meat in the meat product. Specific primers and probes were designed according to the D-loop gene sequence of the pork mitochondria（mtDNA），then the concentration of primers and reaction condition were optimized to establish the best system eventually. The results showed that the amplification system had good specificity for the pork-derived component as the detection limit of the single pork-derived component reached 1 ng/μL，while detection ratio of the pork meat component was 10%（equivalent to 10 ng/μL）. Nucleic acid test strips could quickly get the amplification products，and the result of test strips was as same as that of gel electrophoresis system. The CPA- nucleic acid test strip method established by this study was a specific and efficient method to detect the component of pork meat，which could provide a new effective approach for the quality control of meat products.

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