School of Applied Sciences, Health Innovations Research Institute, RMIT University, Melbourne, Victoria 3000, Australia.
J Sci Food Agric. 2011 May;91(7):1310-5. doi: 10.1002/jsfa.4319. Epub 2011 Feb 18.
Food adulteration remains a major global concern. DNA fingerprinting has several advantages over chemical and morphological identification techniques. DNA microarray-based fingerprinting techniques have not been used previously to detect adulteration involving dried commercial samples of closely related species. Here we report amplification of low-level DNA obtained from dried commercial ginseng samples using the Qiagen REPLI-g Kit. Further, we used a subtracted diversity array (SDA) to fingerprint the two ginseng species, Panax ginseng and Panax quinquefolius, that are frequently mixed for adulteration.
The two ginseng species were successfully discriminated using SDA. Further, SDA was sensitive enough to detect a deliberate adulteration of 10% P. quinquefolius in P. ginseng. Thirty-nine species-specific features including 30 P. ginseng-specific and nine P. quinquefolius-specific were obtained. This resulted in a feature polymorphism rate of 10.5% from the 376 features used for fingerprinting the two ginseng species. The functional characterization of 14 Panax species-specific features by sequencing revealed one putative ATP synthase, six putative uncharacterized proteins, and two retroelements to be different in these two species.
SDA can be employed to detect adulterations in a broad range of plant samples.
食品掺假仍然是一个全球性的主要问题。与化学和形态识别技术相比，DNA 指纹图谱具有多个优势。以前没有使用基于 DNA 微阵列的指纹图谱技术来检测涉及密切相关物种的干燥商业样品的掺假。在这里，我们报告了使用 Qiagen REPLI-g 试剂盒从干燥的商业人参样品中扩增低水平 DNA 的方法。此外，我们使用差减多样性阵列 (SDA) 对经常混合掺假的两种人参，即 Panax ginseng 和 Panax quinquefolius，进行指纹图谱分析。
使用 SDA 成功地区分了这两种人参。此外，SDA 足够灵敏，能够检测到故意掺入 10%的 P. quinquefolius 到 P. ginseng 中。获得了 39 个物种特异性特征，包括 30 个 P. ginseng 特异性特征和 9 个 P. quinquefolius 特异性特征。这导致用于对两种人参进行指纹图谱分析的 376 个特征中有 10.5%的特征多态性。通过测序对 14 个 Panax 种特异性特征的功能表征揭示了这两个物种中的一个假定 ATP 合酶、六个假定未鉴定的蛋白质和两个逆转录元件不同。
SDA 可用于检测广泛的植物样品中的掺假。
