摘要
目的建立一种快速、准确、灵敏的多重PCR检测方法,用于同时鉴别6种常见食用肉类(牛、羊、鸡、猪、鹅、鸭),并评估其在肉类食品掺假鉴定中的应用价值。方法基于GenBank数据库中6个物种的线粒体全基因组序列,筛选具有种内保守性、种间特异性的DNA序列(牛16S rRNA、羊COX-1、鸡Cytb、猪COX-1、鹅NADH2、鸭16S rRNA),并设计物种特异性引物,以此构建一个可同时鉴别6种常见食用肉类的多重PCR检测体系。对该体系进行种属特异性、灵敏度和重复性研究,并进行模拟混合样品检测。结果成功构建了一个可同时鉴别6种常见食用肉类的多重PCR检测体系,该体系在非目标物种的DNA中均未有效扩增;在DNA模板量为0.0625~2ng/μL时,6个物种的扩增产物均能检见。在鸭肉和牛肉混合比例低至0.5%时,仍能检测到鸭肉成分。结论本研究构建了一个特异性强、灵敏度高、重复性好的多重PCR检测体系,可准确鉴别6种常见食用肉类食品中的动物源性成分,为我国常见食用肉类及肉制品的掺假鉴定提供了一种简单、实用的检测技术。
Objective To establish a rapid,accurate,and sensitive multiplex PCR detection method for the simultaneous identification of the six common edible meats(beef,lamp,chicken,pork,goose,duck),and to evaluate its application value in meat adulteration identification.Methods Based on complete mitochondrial genomic sequences of six species in the GenBank database,DNA sequences(cattle:16S rRNA;sheep:COX-1;chickens:Cytb;pig:COX-1;goose:NADH2;duck:16S rRNA)with intra-species conservation and inter-species specificity were screened,and species-specific primers were designed to construct a multiplex PCR detection system that can simultaneously detect the meat of six common species.The species specificity,sensitivity and reproducibility of the system were studied,and the simulated mixture sample detection was performed.Results This study successfully constructed a multiplex PCR detection system that can detect the meats of six common species simultaneously.The system was not effective in DNA amplification of non-target species.When the DNA template sizes were 0.0625-2 ng/μL,the amplified products of all six species could be detected.The duck component was still detected when the mixing ratio of duck and beef was as low as 0.5%.Conclusion This study constructs and establishes a multiplex PCR detection system with strong specificity,high sensitivity,and good reproducibility.It can accurately identify the components of animal origin in common edible meats and provide a simple and practical method for identifying adulteration of common edible meats and meat products in China.
作者
蒋志伟
夏若成
陶瑞旸
李成涛
Zhi-wei JIANG;Ruo-cheng XIA;Rui-yang TAO;Cheng-tao LI(Department of Forensic Medicine,School of Basic Medical Sciences,Wenzhou Medical University,Wenzhou 325035,Zhejiang Province,China;Shanghai Key Laboratory of Forensic Medicine,Key Laboratory of Forensic Science,Ministry of Justice,Shanghai Forensic Service Platform,Academy of Forensic Science,Shanghai 200063,China)
出处
《法医学杂志》
CAS
CSCD
北大核心
2024年第3期254-260,共7页
Journal of Forensic Medicine
基金
国家自然科学基金资助项目(81930056)
上海市法医学重点实验室资助项目(21DZ2270800)
司法部司法鉴定重点实验室资助项目
上海市司法鉴定专业技术服务平台资助项目。
