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中国精品科技期刊2020
励炯,吴琼,江海,等. 基于毛细管凝胶电泳与DNA条形码技术鉴别可食用动物内脏掺假方法的研究[J]. 食品工业科技,2023,44(15):329−336. doi: 10.13386/j.issn1002-0306.2022090211.
引用本文: 励炯,吴琼,江海,等. 基于毛细管凝胶电泳与DNA条形码技术鉴别可食用动物内脏掺假方法的研究[J]. 食品工业科技,2023,44(15):329−336. doi: 10.13386/j.issn1002-0306.2022090211.
LI Jiong, WU Qiong, JIANG Hai, et al. Identification of Adulterated Animal-derived Ingredients in Edible Animal Viscera Based on Capillary Gel Electrophoresis and DNA Barcoding Techniques[J]. Science and Technology of Food Industry, 2023, 44(15): 329−336. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022090211.
Citation: LI Jiong, WU Qiong, JIANG Hai, et al. Identification of Adulterated Animal-derived Ingredients in Edible Animal Viscera Based on Capillary Gel Electrophoresis and DNA Barcoding Techniques[J]. Science and Technology of Food Industry, 2023, 44(15): 329−336. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022090211.

基于毛细管凝胶电泳与DNA条形码技术鉴别可食用动物内脏掺假方法的研究

Identification of Adulterated Animal-derived Ingredients in Edible Animal Viscera Based on Capillary Gel Electrophoresis and DNA Barcoding Techniques

  • 摘要: 建立并优化了使用基于DNA条形码技术对可食用内脏制品中包括猪、牛、羊、鸡、鸭、鹅、兔7种常见动物源成分进行掺假鉴别的方法。用生理盐水清洗和真空冷冻干燥预处理后的内脏样品,经DNA提取扩增后,扩增产物经毛细管凝胶电泳分析系统进行确认,克隆测序结果提交本地数据库Viscera进行比对,同时筛选出适合7种动物源内脏DNA扩增的通用引物COI-A,优化DNA模板量和退火温度,验证考察了19个可食用内脏掺假模型的最低掺假比例。7种动物的5类内脏的PCR扩增效率均为100%,最佳的DNA模板量和退火温度为2 μL和53 ℃,掺假成分的最低检出比例为5%。本方法灵敏度高,可靠性好,可作为常见可食用动物内脏掺假的有效检测方法。

     

    Abstract: A DNA barcoding method with cytochrome C oxidase subunit I sequence (COI) was developed to identify 7 adulterated animal-derived components (including pig, cattle, sheep, chicken, duck, goose and rabbit) in edible viscera products. Samples were cleaned with physiological saline and pretreated by vacuum freeze drying before DNA extraction and amplification. PCR products were confirmed by capillary gel electrophoresis analysis system, and the cloned sequencing results were submitted to the local database (Viscera) for comparison. The universal primer set COI-A was used for the amplification, and the amount of DNA template and annealing temperature were optimized. Meanwhile, the minimum adulteration percentage of 19 edible viscera adulteration models was validated and examined. Results showed that the 5 viscera sources from 7 animal species can be completely amplified under the above conditions, the optimal DNA template volume and annealing temperature are 2 μL and 53 ℃ respectively, and the minimum detection percentage of adulterated components was 5%. The method is sensitive and reliable, which can be used for the identification of adulterated 7 animal-derived components in the edible viscera products.

     

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