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Direct duplex real-time loop mediated isothermal amplification assay for the simultaneous detection of cow and goat species origin of milk and yogurt products for field use

机译:直接双工实时环路介导的等温扩增测定法,用于同时检测现场使用的牛奶和酸奶产品的牛和山羊种

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摘要

A multiple loop-mediated isothermal amplification (LAMP) method was developed to detect cow and goat milk in the field using a portable fluorescence device. For rapid on-site detection, this duplex LAMP assay was used in combination with direct amplification, without DNA extraction. The cow-and goat-specific LAMP primer sets were designed based on the mitochondrial cytochrome b gene, and showed specificity against 13 other animal species in the reactions. The sensitivity of the duplex LAMP assay for cow and goat was 0.1 and 1 pg, respectively. The detection limit for both target species in milk mixtures was 2%. This assay successfully amplified and identified the two target species in 24 samples of commercial milk and yogurt products, with 30 min sampling-to-result analysis time. Therefore, this direct duplex real-time LAMP assay is useful for on-site simultaneous detection of cow and goat milk in commercial products, a capability needed to confirm accurate labeling.
机译:开发了一种多环介导的等温扩增(LAMP)方法,以使用便携式荧光设备在现场检测牛和山羊奶。为了快速进行现场检测,此双LAMP检测与直接扩增结合使用,无需提取DNA。基于线粒体细胞色素b基因设计了牛和山羊特异性LAMP引物,并在反应中显示出对其他13种动物的特异性。牛和山羊的双重LAMP检测灵敏度分别为0.1 pg和1 pg。牛奶混合物中两种目标物质的检出限均为2%。该测定法成功扩增并鉴定了24种商品奶和酸奶产品样品中的两个目标物种,采样至结果分析时间为30分钟。因此,这种直接双工实时LAMP测定法可用于现场同时检测商业产品中的牛和山羊奶,这是确认准确标记所必需的能力。

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