【摘要】 根据线粒体保守序列选取牛、羊特异性引物,建立无需核酸提取的直接实时环介导等温扩增(LAMP)技术,用于羊乳中掺入牛乳成分的检测。结果表明,所建立的LAMP方法特异性好,对其他物种没有交叉反应及假阳性现象的发生,对牛、羊的检测灵敏度可达0.07 pg/μL和0.7 pg/μL,在掺假样品(即二元混合乳)中直接实时LAMP技术可检测到掺入1%的牛乳成分。通过对10种市售羊乳制品检测显示,直接实时LAMP方法与经DNA提取的实时LAMP方法检测结果一致。因此建立的直接实时LAMP技术无需核酸提取,样品经简单处理即可作为模板直接进行LAMP扩增,简化了样品前处理过程,缩短了检测时间,适用于羊乳制品中牛乳成分的快速检测。更多还原

【Abstract】 According to the mitochondria conserved sequence, bovine and goat specific primers were selected to establish a direct real-time Loop mediated isothermal amplification(LAMP) technique without nucleic acid extraction, which was used to detect the incorporation of cow milk in goat milk. The results showed that the established LAMP method had good specificity, no cross-reactions and false positives for other species, and the detection sensitivity of cow and goat was up to 0.07 pg/μL and 0.7 pg/μL. The detection limit for binary mixed milk was 1%. The detection of ten commercially available goat dairy products showed that the direct real-time LAMP was consistent with the conventional real-time LAMP method using extracted DNA. Therefore, the established direct real-time LAMP technology does not require nucleic acid extraction, and the sample can be used as a template for direct LAMP amplification after simple treatment, which simplifies the sample pretreatment process, shortens the detection time, and is suitable for rapid detection of cow milk components in goat dairy products.更多还原