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題名 | 新型核酸分子等溫擴增技術於中藥材基因體基源鑑定之研究--以常見誤用及混用中藥材之基源為例=Study on Genomic Authentication of Chinese Herbal Medicine Using a Novel Isothermal Nucleic Acid Amplification Technology--Creating an Authentication Model for Mis-identified Medicinal Herb Species and Its Adulterants |
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編 次 | (2-1) |
作者 | 李孟修; | 書刊名 | 中醫藥年報 |
卷期 | 3 2014.12[民103.12] |
頁次 | 頁(12)1-(12)39 |
分類號 | 414.32 |
關鍵詞 | 中藥品質管制; 等溫圈環式核酸擴增法; 誤混用藥材; Quality control in the traditional medicines; Loop-mediated isothermal amplification; Adulterants; |
語文 | 中文(Chinese) |
中文摘要 | 中藥品質管制的目的,在於辨別真偽、參雜及品質的優劣,以確保臨床的療效。而誤用、混用以及偽劣藥材,不僅無助於疾病之治療,甚至會危害國人的健康。因此,如何有效鑒定及管控中藥材,為當前刻不容緩的重要課題。中草藥分子鑒定是運用 DNA分子遺傳標記技術,直接分析中草藥原植物的基因型而非表現型(如型態及化學成分),不受材料來源、環境因素、樣品形態(原品、粉末或片狀)影響,樣品少數即可進行實驗。 本計劃應用等溫圈環式核酸擴增法 (LAMP),以三種常見易誤混用藥材為模式藥材,針對中藥材正品藥材之目標基因如核糖體核基因(例如 ITS)或葉綠體基因組(例如 trnL-trnF IGS)中的 6個特異區域,設計 4種特異引子 (Primer),並在恆溫條件下 (65℃)完成標的基因之核酸擴增 (Amplification)反應,藉以區辨正品及其易誤、混用藥材。本研究成功建立白花蛇舌草 (Hedyotis diffusa)、蒲公英 (Taraxacum formosanum)及粉防己 (Stephania tetrandra)正品藥材與馬兜鈴屬藥材之恆溫圈環式核酸擴增法,此法具有高特異性且可快速的鑑定這些藥材之基源。 運用並建立 LAMP做為植物基因體鑑別的方法,可針對中藥材正品與其混用藥材基因組,設計特定引子,藉由最佳化反應條件,可觀察中藥材正品及混用藥材的 LAMP檢測效果,其靈敏度和特異性皆相當高,此研究為中草藥分子鑒定技術提供了一鑑定種強而有力的新方法。 |
英文摘要 | Quality control plays a vital role in the development of translating the traditional medicines into modern evidence-based therapies. Authentication of traditional Chinese medicine (TCM) is generally recommended for the standardization and quality control of herbal products and herb related investigations. It has been shown that counterfeits and inferior medicinal material reduce TCM quality and cause cases which patients do not recover or even die. Hence, authentication is an essential step for clinical application and accurate experimental studies of TCM. In recent years, the DNA molecular biology method have now become a popular method for the identification and authentication of TCM and developed to be a more effective, accurate, reliable and sensitive technology. Loop-mediated isothermal amplification (LAMP) assay is a potential method that is capable of amplifying DNA with high specificity, rapidity, and efficiency under isothermal conditions. The method is under isothermal conditions (65℃)using four to six specifically designed primers. One key feature of the LAMP is using Bst DNA polymerase instead of Taq DNA polymerase in the reaction. Therefore, the aim of this study is to use LAMP assay for the detection of ITS sequence/ trnL-trnF IGS in two plants of similar species such as H. diffusa and H. corymbosa, Taraxacum formosanum and Ixeris chinensis or other misused Aristolochiaceae plants such as Stephania tetrandra and Aristolochia fungchi. In this study, the LAMP method for rapid and sensitive detecting three herbal plants is successfully established, and the LAMP method will be applied for the detection of adulterants. It might be an alternative method to rapid authenticates the TCM at the market in the future. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。