Zhi-Wei Jiang, Ruo-Cheng Xia, Rui-Yang Tao, Cheng-Tao Li
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The species specificity, sensitivity and reproducibility of the system were studied, and the simulated mixture sample detection was performed.</p><p><strong>Results: </strong>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 0.062 5-2 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%.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":12317,"journal":{"name":"法医学杂志","volume":"40 3","pages":"254-260"},"PeriodicalIF":0.0000,"publicationDate":"2024-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Establishment and Validation of a Multiplex PCR Detection System for the Identification of Six Common Edible Meat Components.\",\"authors\":\"Zhi-Wei Jiang, Ruo-Cheng Xia, Rui-Yang Tao, Cheng-Tao Li\",\"doi\":\"10.12116/j.issn.1004-5619.2023.531002\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objectives: </strong>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.</p><p><strong>Methods: </strong>Based on complete mitochondrial genomic sequences of six species in the GenBank database, DNA sequences (cattle:<i>16S rRNA</i>; sheep:<i>COX-1</i>; chickens:<i>Cytb</i>; pig:<i>COX-1</i>; goose:<i>NADH2</i>; duck:<i>16S rRNA</i>) 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. 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引用次数: 0
摘要
目的建立一种快速、准确、灵敏的多重PCR检测方法,用于同时鉴定6种常见食用肉类(牛、灯、鸡、猪、鹅、鸭),并评价其在肉类掺假鉴定中的应用价值:方法:基于 GenBank 数据库中六种肉类的完整线粒体基因组序列,对其 DNA 序列(牛:16S rRNA;羊:COX-1;鸡:Cytb;猪:COX-1;鹅:NADH2;鸭:16S rRNA)的种内保守性和种间特异性,设计了种间特异性引物,构建了可同时检测 6 种常见肉类的多重 PCR 检测系统。研究了该系统的物种特异性、灵敏度和重现性,并进行了模拟混合样品检测:结果:本研究成功构建了一个可同时检测六种常见肉类的多重 PCR 检测系统。该系统对非目标物种的 DNA 扩增无效。当 DNA 模板大小为 0.062 5-2 ng/μL 时,可检测到所有六个物种的扩增产物。当鸭肉和牛肉的混合比例低至 0.5%时,仍能检测到鸭肉成分:本研究构建并建立了一种特异性强、灵敏度高、重现性好的多重 PCR 检测系统。结论:该研究构建并建立了特异性强、灵敏度高、重现性好的多重 PCR 检测系统,可准确鉴定常见食用肉类中的动物源性成分,为我国常见食用肉类及肉制品的掺假鉴别提供了一种简便实用的方法。
Establishment and Validation of a Multiplex PCR Detection System for the Identification of Six Common Edible Meat Components.
Objectives: 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 0.062 5-2 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%.
Conclusions: 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.