实时荧光PCR检测动物源性基因鉴定地沟油

Identification of hogwash oil through animal-original genes detection with real-time PCR

目的寻找一个多种动物基因组共有的基因片段作为地沟油鉴定的特异性指标,建立准确、特异、快速的地沟油鉴定方法。方法通过多个数据库的比对分析筛选出靶标基因片段,采用实时荧光PCR方法,以常见动植物组织、正常食用植物油以及6份地沟油作为验证样品对新建方法的特异性、灵敏度以及在地沟油鉴定中的适用性进行验证。结果筛选得到的HOXC5基因片段在7种动物组织中均有扩增,而在11种植物组织中均没有扩增;此外,8份正常食用植物油样品中均被鉴定为阴性,而6份地沟油样品均被鉴定为阳性。结论筛选出的基因片段具有动物组织特异性,新建方法能够正确区分正常食用植物油和地沟油样品,其检测灵敏度与使用物种特异性动物基因作为检测靶标的方法相当。

Objective Common gene fragments from multiple animal genomes were selected as specific targets for accurate and quick identification of hogwash oil. Methods Target gene fragment was screened by analyzing multiple databases and validated by real- time PCR （RT- PCR） to testify the specificity, sensitivity and serviceability in samples of common animal and plant tissues, normal vegetable oils and six hogwash oils. Results HOXC5 gene fragment was found amplification in all the 7 animal tissues while none in the 11 plant tissues. All 8 normal vegetable oil samples showed negative, while all 6 hogwash oil samples showed positive by the RT- PCR. Conclusion The screened gene fragment could differentiate normal vegetable oil from hog- wash oil with multiple animal - tissues specificity, and the sensitivity was the same as the previous detection method with species -specific gene from animal.