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CN103740810A 一种猪肠道组织中粘蛋白12荧光定量PCR检测方法 无效 - 驳回

一种猪肠道组织中粘蛋白12荧光定量PCR检测方法
摘要：
本发明涉及一种猪肠道组织中粘蛋白（MUC）12转录本荧光定量PCR检测方法。该方法包括设计特异性荧光定量PCR引物，建立荧光定量PCR反应体系和反应程序，制备含有目的片段MUC12的质粒作为标准质粒绘制标准曲线，最后待测样品的目的基因和内参基因GAPDH同时扩增，根据相对定量△△CT方法检测基因的表达量。本发明由于利用了荧光定量PCR具有灵敏度高，特异性强，无污染且实时、准确、快速的特点，从而填补了该基因在猪肠道组织中检测方法的空白，为精确检测猪肠道部位的MUC12表达水平，从而辅助评价动物肠道粘膜屏障及其免疫状态提供了一个实用方法。
摘要（英）：

The invention relates to a fluorescent quantitative PCR detection method of mucoprotein (MUC) 12 in porcine intestinal tissues. The method comprises the following steps: designing a specific fluorescent quantitative PCR primer, establishing a fluorescent quantitative PCR reaction system and a reaction program, preparing a plasmid containing a target fragment MUC12 as a standard plasmid to draft a standard curve, simultaneously amplifying the target gene and internal control gene GAPDH in a sample for measurement, and detecting the expression level of the gene through a quantitative delta delta CT method. The fluorescent quantitative PCR detection method utilizing fluorescent quantitative PCR has the characteristics of high sensitivity, strong specificity, no pollution, real time, accuracy and rapidness, so the fluorescent quantitative PCR detection method fills up a gap in the detection method of the gene in the porcine intestinal tissues, and is a practical method for accurately detecting the MUC12 expression level of the porcine intestinal part in order to realize the auxiliary evaluation of the mucosal barrier of animal intestines and the immune states thereof.

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