基于DNA条形码和散射元基因组学的草药产品Gurigumu-7中生物成分的物种鉴定

Miaojie Wei, Yu Tian, Erhuan Zang, Battseren Tsambaa, Jin-xin Liu, Linchun Shi, Almaz Borjigidai
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引用次数: 0

摘要

准确鉴定混合物中的物种组成是一项重大挑战,尤其是在由多种物种组成的加工混合物中,如食品和药品中的混合物。因此,我们尝试利用枪式代谢编码技术来解决这一问题。在这项研究中,我们首先利用蒙古化合物制剂 Gurigumu-7 (G-7) 的两个模拟样本建立了该方法,然后将其应用于 3 种药品和 12 种医院制剂。通过枪式元基因组测序,共获得了 119.72 Gb 的原始数据集。通过结合 ITS2、matK 和 rbcL,可以检测到 G-7 处方中规定的所有标记生物成分,尽管某些物种可能无法在所有样本中检测到。除 YN02 和 YN12 外,在所有药品和医院样本中都能找到普遍的 Akebia quinata 替代品。在 YN02 样品中只发现了 Akebia quinata 的毒性替代品--马兜铃属 manshuriensis。为了进一步证实这一结果,我们在 YN02 中使用 HPLC 和 TaqMan 探针进行实时 PCR 验证。结果表明,通过高效液相色谱法在 YN02 中检测到了马兜铃酸 A(AAA),并通过 qPCR 和 TaqMan 探针在 YN02 中验证了马兜铃属植物的 ITS2 序列。这项研究证实了枪式代谢编码能有效鉴定蒙药复方制剂 G-7 中的生物成分。该研究还证明了该方法可用作含有多种植物的混合物的通用鉴定技术。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Species identification of biological ingredients in herbal product, Gurigumu-7, based on DNA barcoding and shotgun metagenomics
Accurate identification the species composition in mixtures poses a significant challenge, especially in processed mixtures comprising multiple species, such as those found in food and pharmaceuticals. Therefore, we have attempted to utilize shotgun metabarcoding technology to tackle this issue. In this study, the method was initially established using two mock samples of the Mongolian compound preparation Gurigumu-7 (G-7), which was then applied to three pharmaceutical products and 12 hospital-made preparations. A total of 119.72 Gb of raw data sets were obtained through shotgun metagenomic sequencing. By combining ITS2, matK, and rbcL, all the labeled bio-ingredients specified in the G-7 prescription can be detected, although some species may not be detectable in all samples. The prevalent substitution of Akebia quinata can be found in all the pharmaceutical and hospital samples, except for YN02 and YN12. The toxic alternative to Akebia quinata, Aristolochia manshuriensis, was exclusively identified in the YN02 sample. To further confirm this result, we validated it in YN02 using HPLC and real-time PCR with TaqMan probes. The results showed that aristolochic acid A (AAA) was detected in YN02 using HPLC, and the ITS2 sequence of Aristolochia manshuriensis has been validated in YN02 through qPCR and the use of a TaqMan probe. This study confirms that shotgun metabarcoding can effectively identify the biological components in Mongolian medicine compound preparation G-7. It also demonstrates the method’s potential to be utilized as a general identification technique for mixtures containing a variety of plants.
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