摘要
通过基于荧光定量PCR方法鉴定粗胰酶种属来源,用于胰激肽原酶和弹性酶药品生产企业的原料验收和质量控制。该方法采用荧光定量PCR法,针对猪、牛、绵羊、山羊基因组特有的基因序列设计引物与探针,采用聚合酶链式反应对目标基因进行扩增,利用扩增曲线的Ct值绘制标准曲线,通过未知样品的Ct值对未知样品初始模板中的基因进行鉴定。结果显示,该方法在实验室条件下具有较好的专属性和耐用性,可保证被检基因的真实性、原始性、完整性。
The quality control method of crude pancreatin species origin based on Fluorescence quantitative PCR was applied in acceptance and quality control of raw materials for pharmaceutical manufacturers of Pancreatic Kininogenase and Elastase. Real-time PCR was used to design primers and probes for genome-specific gene sequences of Porcine,Bovine,Ovine and Caprine. Polymerase chain reaction was used to amplify the target gene. The standard curve was drawn by the Ct value of the amplification curve,and the genes in the initial template of unknown samples were identified by the Ct value of unknown samples. Results showed that this method had sufficient specificity and durability under the laboratory conditions,and authenticity,aboriginality and integrity of the tested genes could be ensured.
引文
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