Abstract
[Objective] To establish a detection method for egg components in food using microfluidic digital polymerase chain reaction (PCR) technology. [Methods] Specific primers and fluorescent probes are designed based on the mitochondrial gene nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1). Then, the reaction system and amplification conditions are optimized, and specificity and sensitivity are evaluated before being applied to actual food detection. [Results] The established method can specifically detect egg components, while no cross-reactivity is observed with other common poultry egg components, such as duck and goose eggs. The optimal annealing temperature is 56 ℃, and the optimal concentrations of primers and probes are 800 nmol/L and 200 nmol/L, respectively. Under these conditions, the limit of detection (LOD) for microfluidic digital PCR is 5.30 copies/μL. Furthermore, the method is applied to detect 20 batches of food samples, yielding consistent results with the sample labels. [Conclusion] Characterized by anti-interference, high sensitivity, rapidity, and simplicity, the established microfluidic digital PCR method can be applied to detect trace egg components in food.
Publication Date
9-20-2026
First Page
49
Last Page
54
DOI
10.13652/j.spjx.1003.5788.2025.80797
Recommended Citation
Ben, Li; Yiming, Zhuo; Jijun, Qin; and Lixia, Yang
(2026)
"Detection of egg components in food by microfluidic digital PCR,"
Food and Machinery: Vol. 42:
Iss.
8, Article 7.
DOI: 10.13652/j.spjx.1003.5788.2025.80797
Available at:
https://www.ifoodmm.cn/journal/vol42/iss8/7
References
[1] Xu J Y,Ye Y L,Ji J,et al.Advances on the rapid and multiplex detection methods of food allergens [J].Crit Rev Food Sci Nutr,2022,62(25):6 887-6 907.
[2] Ismail A M,Sani M S A,Azid A,et al.Food forensics on gelatine source via ultra-high-performance liquid chromatography diode-array detector and principal component analysis [J].SN Applied Sciences,2021,3(1):79.
[3] 潘志文,黄婉锋,钱鑫,等.基于超高效液相色谱串联质谱法特征肽检测技术的阿胶鉴别与6种动物异源性成分检查 [J].中国药业,2024,33(11):79-83.Pan Z W,Huang W F,Qian X,et al.Identification of colla corii asini and examination of six animal heterologous components based on UPLC-MS/MS characteristic peptide detection technology [J].China Pharmaceuticals,2024,33(11):79-83.
[4] Liu C Q,Su M N,Champ V D,et al.Validation procedures for quantification of food allergens by enzyme-linked immunosorbent assay (ELISA) [J].Methods in Molecular Biology,2024,2 717:285-304.
[5] Adenuga B M,Biltes R,Villa C,et al.Unravelling red deer (Cervus elaphus) meat adulteration in gourmet foods by quantitative real-time PCR [J].Food Control,2025,168:110872.
[6] 蒋志伟,夏若成,陶瑞旸,等.鉴别6种常见食用肉类的多重PCR检测体系的构建及验证 [J].法医学杂志,2024,40(3):254-260.Jiang Z W,Xia R C,Tao R Y,et al.Establishment and validation of a multiplex PCR detection system for the identification of six common edible meat components [J].Journal of Forensic Medicine,2024,40(3):254-260.
[7] 金萍,丁洪流,张敏,等.双重实时荧光PCR法测定食品中大豆、小麦过敏原 [J].食品与机械,2022,38(9):59-63,102.Jin P,Ding H L,Zhang M,et al.Determination of soybean and wheat allergens in food by dual real-time fluorescence PCR [J].Food & Machinery,2022,38(9):59-63,102.
[8] 易姿,杨丽霞,钟菲菲,等.食品中植物源性过敏原成分的DNA条形码鉴定 [J].食品与机械,2025,41(10):20-26.Yi Z,Yang L X,Zhong F F,et al.Component identification of plant-derived allergens in food based on DNA barcoding [J].Food & Machinery,2025,41(10):20-26.
[9] 路宇轩,翟睿,吴帆,等.肉类掺假检测方法及研究进展 [J].计量学报,2023,44(6):1 000-1 008.Lu Y X,Zhai R,Wu F,et al.Research progress on detection technology of meat adulteration [J].Acta Metrologica Sinica,2023,44(6):1 000-1 008.
[10] Francis O L,Wang K Y,Kim E H,et al.Common food allergens and cross-reactivity [J].Journal of Food Allergy,2020,2(1):17-21.
[11] Nigolian H,Caubet J C,Coattrenec Y.Egg allergy in children and adults [J].Revue Medicale Suisse,2021,17(733):675-679.
[12] 杨文华,叶惠玲,王辉,等.鸡蛋过敏原及其脱敏技术研究进展[J].食品科学,2025,46(10):333-345.Yang W H,Ye H L,Wang H,et al.Research advances in hen's egg allergen and techniques for reducing its allergenicity [J].Food Science,2025,46(10):333-345.
[13] Wang H,Chen L G,Sun L N.Digital microfluidics:a promising technique for biochemical applications [J].Frontiers of Mechanical Engineering,2017,12(4):510-525.
[14] 冯秀晶,易红梅,任星旭,等.数字PCR技术及其在检测领域的应用 [J].遗传,2020,42(4):363-373.Feng X J,Yi H M,Ren X X,et al.Digital PCR and its application in biological detection [J].Hereditas,2020,42(4):363-373.
[15] 黄瑾,梁涛波,许恒毅.数字PCR在生物学检测中应用的研究进展 [J].生命科学,2021,33(2):255-264.Huang J,Liang T B,Xu H Y.Research progress of application of digital PCR in biological detection [J].Chinese Bulletin of Life Sciences,2021,33(2):255-264.
[16] 李恩静,王丹,李龙,等.基于微滴数字PCR技术检测致敏原芒果成分 [J].食品与机械,2024,40(9):50-55,115.Li E J,Wang D,Li L,et al.Detection of allergen mango components by using droplet digital PCR technology [J].Food & Machinery,2024,40(9):50-55,115.
[17] 闫邦奇,水克娟,黎财慧,等.转基因玉米MON 87411双重微滴式数字PCR定量检测方法的建立 [J].现代食品科技,2025,41(3):378-386.Yan B Q,Shui K J,Li C H,et al.Establishment of a quantitative duplex droplet digital PCR detection method for genetically modified maize MON 87411 [J].Modern Food Science and Technology,2025,41(3):378-386.
[18] 张雪,马丽侠,叶德萍,等.即食果蔬制品中致泻大肠埃希氏菌检测微滴数字PCR系统的建立 [J].食品工业科技,2025,46(21):357-365.Zhang X,Ma L X,Ye D P,et al.Establishment of a droplet digital PCR system for the detection of diarrheagenic Escherichia coli in ready-to-eat fruit and vegetable products [J].Science and Technology of Food Industry,2025,46(21):357-365.
[19] 陈晨.微滴式数字PCR技术用于牛羊肉掺假定量检测方法的研究 [D].保定:河北农业大学,2018:17.Chen C.Research on microbial digital PCR quantitative detection techniques of adulteration of beef and sheep [D].Baoding:Hebei Agricultural University,2018:17.
[20] 杨华,汪小福,肖英平,等.牛肉及其制品中掺入鸡肉、鸭肉和猪肉的多重数字PCR快速检测方法研究 [J].浙江农业学报,2017,29(6):994-1 000.Yang H,Wang X F,Xiao Y P,et al.Rapid detection of chicken,duck and pork blending in beef products by multiple digital PCR [J].Acta Agriculturae Zhejiangensis,2017,29(6):994-1 000.
[21] 国家市场监督管理总局,国家标准化管理委员会.GB/T 38164—2019 常见畜禽动物源性成分检测方法 实时荧光PCR法[S].北京:中国标准出版社,2019.State Administration for Market Regulation,Standardization Administration of the People's Republic of China.GB/T 38164—2019 Identification of animal ingredient from common livestock and poultry—real-time PCR [S].Beijing:Standards Press of China,2019.
[22] 中华人民共和国商务部.SB/T 10923—2012 肉及肉制品中动物源性成分的测定 实时荧光PCR法[S].北京:中国标准出版社,2013.Ministry of Commerce of the People's Republic of China.SB/T 10923—2012 Identification of animal derived materials in meat and meat products real-time PCR method [S].Beijing:Standards Press of China,2013.
[23] 国家市场监督管理总局.BJS 202309 鸭血中鸭鸡鹅源性成分的测定 [S].北京:中国标准出版社,2023.State Administration for Market Regulation.BJS 202309 Determination of duck,chicken and goose-derived components in duck blood [S].Beijing:Standards Press of China,2023.
[24] Group D,Huggett J F.The digital MIQE guidelines update:minimum information for publication of quantitative digital PCR experiments for 2020 [J].Clinical Chemistry,2020,66(8):1 012-1 029.
[25] Walia G K,Dhillon G K.DNA barcoding and molecular phylogeny of genus Pseudagrion (Odonata:Coenagrionidae) based on mitochondrial COI,ND1 and 16S rRNA genes [J].International Journal of Tropical Insect Science,2024,44(4):1 567-1 590.
[26] Codex Alimentarius Commission,CAC.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].Rome:Codex Alimentarius Commission,2010.
