YANG Yao, SI Nengwu, YAN Yuao, et al. Detection of Pork-Derived Ingredients Based on Real-time Quantitative PCR[J]. Science and Technology of Food Industry, 2022, 43(3): 268−274. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021050232.
Citation: YANG Yao, SI Nengwu, YAN Yuao, et al. Detection of Pork-Derived Ingredients Based on Real-time Quantitative PCR[J]. Science and Technology of Food Industry, 2022, 43(3): 268−274. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021050232.

Detection of Pork-Derived Ingredients Based on Real-time Quantitative PCR

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  • Received Date: May 26, 2021
  • Available Online: December 02, 2021
  • In order to accurately and reliably quantify pork-derived ingredients in meat products, single-copy gene (carcinoembryonic antigen-related cell adhesion molecule 2-like, CACA) in pig cell nucleus was screened by bioinformatics methods, with CACA gene as the amplification target, specific primers and TaqMan probes were designed, and a TaqMan real-time quantitative PCR method for detecting pork ingredients was established. The experimental results showed that the method had good extraspecies specificity and intraspecies stability. There was a good linear relationship between the template DNA and the amplified Ct value within the DNA concentration range from 20 to 0.032 ng/µL. The standard curve was y=−3.508x+28.636, the coefficient of determination (R2) value was 0.997. Under the condition of relative standard deviation ≤25%, it could accurately detect DNA samples with pork DNA content as low as 0.1%, and meat products with pork content as low as 5.0%. Through testing of commercially available samples, it was found that somemeat products were mixed with pork but were not labeled. And some meat products, pork was not listed as the main ingredient but in fact pork was the main ingredient. The real-time quantitative PCR method established in this study could be used to accurately and reliably detect pork ingredients in meat products. It has important reference value for accurately identifying adulteration of meat products and quantifying the ingredients of meat products.
  • [1]
    ABBOTS E J, COLES B. Horsemeat-gate: The discursive production of a neoliberal food scandal[J]. Food Culture & Society an International Journal of Multidisciplinary Research,2013,16(4):535−550.
    [2]
    高敬, 魏迪, 张癸荣, 等. 常见肉类鉴别技术研究进展[J]. 食品科学,2014,35(11):356−360. [GAO J, WEI D, ZHANG G R, et al. Recent progress on commonly used techniques for identification of meat species[J]. Food Science,2014,35(11):356−360. doi: 10.7506/spkx1002-6630-201411068
    [3]
    BARNARD C, O'CONNOR N. Runners and riders: The horsemeat scandal, eu law and multi-level enforcement[J]. The Cambridge Law Journal,2017,76(1):116−144. doi: 10.1017/S000819731700006X
    [4]
    张玉华, 孟一, 姜沛宏, 等. 近红外技术对不同动物来源肉掺假的检测[J]. 食品工业科技,2015,36(3):316−319,334. [ZHANG Y H, MENG Y, JIANG P H, et al. Detection of adulteration of animal meats from different sources by near infrared technology[J]. Science and Technology of Food Industry,2015,36(3):316−319,334.
    [5]
    QAMAR Z, MOHAMMAD A, KHAIRIL M N F, et al. Current analytical methods for porcine identification in meat and meat products[J]. Food Chemistry,2020,324(1):126664−126693.
    [6]
    任君安, 黄文胜, 葛毅强, 等. 肉制品真伪鉴别技术研究进展[J]. 食品科学,2016,37(1):247−257. [REN J A, HUANG W S, GE Y Q, et al. Progress in meat adulteration detection techniques[J]. Food Science,2016,37(1):247−257. doi: 10.7506/spkx1002-6630-201601043
    [7]
    BALLIN N Z. Authentication of meat and meat products[J]. Meat Science,2010,86(3):577−587. doi: 10.1016/j.meatsci.2010.06.001
    [8]
    中国标准化研究院, 四川华汉三创生物科技有限公司, 山东省食品药品检验研究院, 等. GB/T 35917-2018常见动物源性成分快速测定 膜芯片法[S]. 北京: 中华人民共和国国家质量监督检验检疫总局, 中国国家标准化管理委员会, 2018.

    CNIS, Sichuan Huahan Trio Biotech, Shandong Institute for Food and Drug Control, et al. GB/T 35917-2018 Rapid detection for animal-derived materials—membrane-based array method[S]. Beijing: General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, Standardization Administration of the People’s Republic of China, 2018.
    [9]
    CAMMA C, DI DOMENICO M, MONACO F. Development and validation of fast real-time PCR assays for species identification in raw and cooked meat mixtures[J]. Food Control,2012,23(2):400−404. doi: 10.1016/j.foodcont.2011.08.007
    [10]
    SAFDAR M, JUNEJO Y. Development and validation of fast duplex real-time PCR assays based on SYBER Green florescence for detection of bovine and poultry origins in feedstuffs[J]. Food Chemistry,2015,173:660−664. doi: 10.1016/j.foodchem.2014.10.088
    [11]
    KIM M, YOO I, LEE S Y, et al. Quantitative detection of pork in commercial meat products by Taqman(r) real-time PCR assay targeting the mitochondrial D-loop region[J]. Food Chemistry,2016,210:102−106. doi: 10.1016/j.foodchem.2016.04.084
    [12]
    HAIDER N, NABULSI I, AL-SAFADI B. Identification of meat species by PCR-RFLP of the mitochondrial COI gene[J]. Meat Science,2012,90(2):490−493. doi: 10.1016/j.meatsci.2011.09.013
    [13]
    AMARAL J S, SANTOS C G, MELO V S, et al. Identification of duck, partridge, pheasant, quail, chicken and turkey meats by species-specific PCR assays to assess the authenticity of traditional game meat alheira sausages[J]. Food Control,2015,47:190−195. doi: 10.1016/j.foodcont.2014.07.009
    [14]
    王婷. 鸡线粒体DNA拷贝数差异研究[D]. 郑州: 河南农业大学, 2017.

    WANG T. Study on the variation of mitochondrial DNA copy number in chicken[D]. Zhengzhou: Henan Agricultural University, 2017.
    [15]
    彭媛媛, 武煊, 陶晓奇. 实时荧光PCR技术定量检测肉类掺假的研究进展[J]. 食品与发酵工业,2019,45(15):279−287. [PENG Y Y, WU X, TAO X Q. Quantitative detection of meat adulteration by real-time fluorescent PCR[J]. Food and Fermentation Industries,2019,45(15):279−287.
    [16]
    BALLIN N Z, VOGENSEN F K, KARLSSON A H. Species determination–can we detect and quantify meat adulteration?[J]. Meat Science,2009,83(2):165−174. doi: 10.1016/j.meatsci.2009.06.003
    [17]
    纪艺, 徐晓丽, 姜媛媛, 等. 基于数字PCR的不同品种鸭组织中线粒体与核DNA拷贝数差异研究[J]. 生物技术通报,2020,36(5):86−91. [JI Y, XU X L, JIANG Y Y, et al. Copy number variations of mitochondrial DNA and genomic DNA from different tissues of duck based on digital PCR[J]. Biotechnology Bulletin,2020,36(5):86−91.
    [18]
    FLOREN C, WIEDEMANN I, BRENIG B, et al. Species identification and quantification in meat and meat products using droplet digital PCR (ddpcr)[J]. Food Chemistry,2015,173:1054−1058. doi: 10.1016/j.foodchem.2014.10.138
    [19]
    陈晨, 张岩, 李永波, 等. 微滴式数字PCR对肉制品中羊肉和猪肉定量分析[J]. 现代食品科技,2018,34(1):221−226,194. [CHEN C, ZHANG Y, LI Y B, et al. Quantitative analysis of mutton and pork in meat products by micro digital PCR[J]. Modern Food Science and Technology,2018,34(1):221−226,194.
    [20]
    支琴, 郭金超, 龚强. 肉制品中动物源性成分DNA检测方法的研究进展[J]. 食品安全质量检测学报,2018,9(16):4204−4211. [ZHI Q, GUO J C, GONG Q. Advance in DNA analytical techniques for ingredients of animal origin in meat products[J]. Journal of Food Safety & Quality,2018,9(16):4204−4211. doi: 10.3969/j.issn.2095-0381.2018.16.004
    [21]
    汪永庆, 王新国, 徐来祥, 等. 一种动物基因组DNA提取方法的改进[J]. 动物学杂志,2001,36(1):27−29. [WANG Y Q, WANG X G, XU L X, et al. A new rapid method for extraction of high quality of genomic DNA from animal tissues[J]. Chinese Journal of Zoology,2001,36(1):27−29. doi: 10.3969/j.issn.0250-3263.2001.01.007
    [22]
    LOPEZ-ANDREO M, LUGO L, GARRIDO-PERTIERRA A, et al. Identification and quantitation of species in complex DNA mixtures by real-time polymerase chain reaction[J]. Anal Biochem,2005,339(1):73−82. doi: 10.1016/j.ab.2004.11.045
    [23]
    FAO, WHO. CAC/GL 74-2010 guidelines on performance criteria and validation of methods for detection, identification and quantification of specific DNA sequences and specific proteins in foods[S]. Codex Alimentarius, 2010.
    [24]
    REN J, DENG T, HUANG W, et al. A digital PCR method for identifying and quantifying adulteration of meat species in raw and processed food[J]. Plos One,2017,12(3):e0173567. doi: 10.1371/journal.pone.0173567
    [25]
    WANG W, FU M, ZHANG Q, et al. A novel quantitative real-time PCR method for the detection of mammalian and poultry species based on a shared single-copy nuclear DNA sequence[J]. Food Chemistry,2021,341(2):128170−128179.
    [26]
    张秀平, 苗丽. 实时荧光PCR定量测定肉制品中羊源性成分[J]. 食品工业科技,2019,40(7):119−123,130. [ZHANG X P, MIAO L. Quantitative detection of ovine-derived materials in meat products by real-time fluorescent PCR[J]. Science and Technology of Food Industry,2019,40(7):119−123,130.
    [27]
    WU Q, XIANG S, WANG W, et al. Species identification of fox-, mink-, dog-, and rabbit-derived ingredients by multiplex PCR and real-time PCR assay[J]. Applied Biochemistry and Biotechnology,2018,185(1):1−12. doi: 10.1007/s12010-017-2621-2
    [28]
    唐廷廷. 羊肉中鸡源性成分实时荧光定量PCR量化检测技术研究[D]. 雅安: 四川农业大学, 2018.

    TANG T T. Quantitative detection of chicken-derived components in mutton by real-time quantitative PCR[D]. Yaan: Sichuan Agricultural University, 2018.
    [29]
    CAI Y C, LI X, LV R, et al. Quantitative analysis of pork and chicken products by droplet digital PCR[J]. Biomed Research International,2014,2014:810209−810214.
    [30]
    张艳, 井汇源, 孙彦婷, 等. 数字PCR精准定量方法研究进展[J]. 中国草食动物科学,2020,40(6):55−59,80. [ZHANG Y, JING H Y, SUN Y T, et al. Research progress on precise quantitative methods of digital PCR[J]. China Herbivore Science,2020,40(6):55−59,80. doi: 10.3969/j.issn.2095-3887.2020.06.013
    [31]
    李婷婷. 基于DNA检测的羊肉掺假鉴别技术研究[D]. 北京: 中国农业科学院, 2019.

    LI T T. Development of DNA-base identification technologies for mutton adulteration[D]. Beijing: Chinese Academy of Agricultural Sciences, 2019.
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