不同核酸提取方法用于检测明胶动物源性成分的比较分析  

Comparative Analysis of Different Nucleic Acid Extraction Methods for Detecting Animal-Derived Components in Gelatin

作　　者：王晶[1,2] 杨彤 芦云 王芳[1,2] WANG Jing;YANG Tong;LU Yun;WANG Fang(Chinese Academy of Inspection and Quarantine Comprehensive Test Center,Beijing 100123,China;CAIQTEST(Beijing)Co.,Ltd.,Beijing 100123,China)

机构地区：[1]中国检验检疫科学研究院,北京100123 [2]中检科(北京)测试技术有限公司,北京100123

出　　处：《食品科技》2024年第3期312-319,共8页Food Science and Technology

基　　金：检科测试集团基本科研业务费项目(CAIQTEST2022004)。

摘　　要：为筛选适用于食用明胶核酸提取的方法,用标准推荐方法、改良十六烷基三甲基溴化铵(Cetyltrimethyl ammonium bromide,CTAB)法、QIAGEN食品基因组脱氧核糖核酸(Deoxyribonucleic acid,DNA)提取试剂盒法和TIANGEN深加工食品因组DNA提取试剂盒法这4种方法提取牛皮明胶、牛骨明胶、猪皮明胶和鱼皮明胶的DNA。通过所提DNA纯度、浓度和物种来源荧光(Polymerase chain reaction,PCR)的扩增效率比较各种方法的提取效果。结果表明,标准推荐方法提取的DNA纯度较低,蛋白污染严重,荧光PCR扩增成功率低。TIANGEN试剂盒法提取DNA纯度高,但仅对皮来源明胶有较好的检测效果,适用范围有限。改良CTAB法和QIAGEN试剂盒法提取DNA纯度高、杂质少,对荧光PCR的抑制小。除鱼皮明胶外,所有样品均检测到了相应的动物源性,可用于大多数明胶样品的DNA提取。In order to select a suitable method for extracting nucleic acids from edible gelatin,the deoxyribonucleic acid(DNA)from bovine skin gelatin,bovine bone gelatin,pig skin gelatin and fish skin gelatin were extracted by four methods:standard recommended method,improved cetyltrimethyl ammonium bromide(CTAB)method,QIAGEN food genome DNA extraction kit method and TIANGEN deeply processed food DNA extraction kit method.The extraction effects of different methods were compared based on DNA purity,concentration and amplification efficiency of fluorescent polymerase chain reaction(PCR).The results showed that the DNA extracted by the standard recommended method had low purity,severe protein contamination,and low success rate of fluorescent PCR amplification.The DNA extracted by TIANGEN kit method exhibited high purity,but it only demonstrated good detection effect on skin source gelatin,with limited scope of application.The improved CTAB method and QIAGEN kit extracted DNA with high purity,less impurities and little inhibition of fluorescent PCR.The corresponding animal derived was detected in all samples except fish skin gelatin,and can be used for DNA extraction of most gelatin samples.

关 键 词：核酸提取 明胶 动物源性 检测 

分 类 号：TS202.3[轻工技术与工程—食品科学]