于智勇, 丁鸽, 丁小余, 褚必海, 钱亮, 顾笋. 猪基源的肝素粗品及混淆品的AS-PCR及ARMS分子检测J. 药学学报, 2008, 43(5): 535-541.
引用本文: 于智勇, 丁鸽, 丁小余, 褚必海, 钱亮, 顾笋. 猪基源的肝素粗品及混淆品的AS-PCR及ARMS分子检测J. 药学学报, 2008, 43(5): 535-541.
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YU Zhi-yong DING Ge, DING Xiao-yu, CHU Bi-hai, QIAN Liang, GU Sun, . Molecular identification of heparin from pigs by allele- specific PCR (AS-PCR) and amplification refractory mutation system (ARMS)J. Acta Pharmaceutica Sinica, 2008, 43(5): 535-541.
Citation: YU Zhi-yong DING Ge, DING Xiao-yu, CHU Bi-hai, QIAN Liang, GU Sun, . Molecular identification of heparin from pigs by allele- specific PCR (AS-PCR) and amplification refractory mutation system (ARMS)J. Acta Pharmaceutica Sinica, 2008, 43(5): 535-541.
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猪基源的肝素粗品及混淆品的AS-PCR及ARMS分子检测

Molecular identification of heparin from pigs by allele- specific PCR (AS-PCR) and amplification refractory mutation system (ARMS)

    摘要
  • 摘要: 本文建立了一种简便、准确的鉴别猪基源的肝素粗品及其混淆品的DNA分子鉴定方法。在对家猪、野猪及其他7种动物(均用于加工猪肝素粗品的混淆品)的mtDNA D-loop区进行序列分析的基础上，设计了专门用于鉴别猪基源肝素钠的AS-PCR(allele-specific PCR)引物及ARMS(amplification refractory mutation system)引物，对家猪、野猪及其他7种动物来源的肝素、肌肉或血液共49个样品的基源进行了分子检测。结果表明：AS-PCR及ARMS方法均可用于猪基源肝素粗品的快速鉴别。AS-PCR鉴别时的复性温度为54～56 ℃，ARMS引物鉴别时的复性温度更宽，为52～58 ℃。在用两种引物对肝素样品进行鉴别时，仅猪来源的肝素DNA模板能扩增得到约170 bp的扩增条带，而其他动物来源的肝素DNA模板在同样条件下无扩增产物。

     

    Abstract: The aim of this study is to develop a convenient and effective method for the identification of heparin from pigs (include Sus scrofa domestica Brisson and Sus scrofa riukiuanus). Based on sequences of D-loop region of pigs and the other animals, two pairs of highly specific primers were designed for distinguishing heparin of pigs from other animals. The primers were employed to amplify D-loop region of DNA templates extracted from pig and seven other animal species that amounted to 49 samples. AS-PCR (allele-specific PCR) and ARMS (amplification refractory mutation system) were all suitable for fast identification of heparin from pig with anneal temperature at 54-56 ℃ in AS-PCR and with wider anneal temperature in ARMS，at 52-58 ℃. An about 170 bp DNA fragments were amplified from separately pigs and whereas no DNA fragment was amplified from other samples under the same reaction condition.

     

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