动物产品中牛、羊源性成分多重PCR检测方法的建立

Development of multiplex PCR detection method for bovine, sheep and goat derived material in animal products

以肉骨粉、鱼粉、猪肉干和鱼肉干为研究对象,异硫氰酸胍法提取总DNA,18S rDNA片段的扩增结果表明提取到的DNA中不存在抑制PCR的物质。应用梯度PCR技术对牛、羊源性成分检测的退火温度进行了优化,在单一PCR检测技术的基础上分别进行了18S rDNA片段和牛、羊源性成分的多重PCR分析,得到了预期的结果。试验表明,本文建立的多重PCR方法具有快速、简便、准确等特点,对动物产品牛、羊源性成分检测具有重要意义。

The total DNAs of meat and bone meal, fish meal, dry pork and dry meat of fish were extracted by guanidine thiocyanate method. The detection results of 18S ribosomal DNA fragment showed that there was no substance restraining polymerase chain reaction (PCR) in the DNAs. The anneal temperature in the detection of bovine, sheep and goat derived materials were optimized by gradient PCR. The multiplex PCR results of 18S ribosomal DNA fragment and bovine, sheep and goat derived material were obstained as expected. The experiment suggested that the multiplex PCR method set up in this paper be quick, simple and practical, and might play an important role on the detection of bovine, sheep and goat derived materials in the animal products.