【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。更多还原

【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.更多还原