基于回折环引物加速LAMP与激烈火球菌Argonaute蛋白的一锅式DNA快速生物传感平台用于肉制品可视化鉴定

丁盛; 李静; 杨明龙; 宋雨佳; 牛翔科; 李美; 罗耀

doi:10.1631/jzus.B2500745

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基于回折环引物加速LAMP与激烈火球菌Argonaute蛋白的一锅式DNA快速生物传感平台用于肉制品可视化鉴定

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Research Article | Updated：2026-07-29

基于回折环引物加速LAMP与激烈火球菌Argonaute蛋白的一锅式DNA快速生物传感平台用于肉制品可视化鉴定
DNA-based one-pot rapid biosensing platform integrating turn-back loop primer-accelerated LAMP and Pyrococcus furiosus Argonaute for meat visual authentication
浙江大学学报（英文版）（B辑：生物医学和生物技术） 2026年27卷第7期页码：775-787
Affiliations：

1.Clinical Medical College & Affiliated Hospital, Chengdu University, Chengdu 610106, China
2.Department of Laboratory Medicine, Sichuan Medical Laboratory Clinical Medicine Research Center, West China Hospital, Sichuan University, Chengdu 610041, China
Funds：

the Sichuan Science and Technology Program(2024YFFK0280);the Natural Science Foundation of Sichuan Province(2024NSFSC0657);the National Natural Science Foundation of China(82402725);the Key Research Project of Clinical Medical College & Affiliated Hospital of Chengdu University(Y202403);the 2024 Undergraduate Innovation Training Program Incubation and Cultivation Project(S202411079085)
DOI：10.1631/jzus.B2500745
中图分类号：
CSTR：
收稿：2025-11-18，

修回：2026-02-17，

网络首发：2026-07-29，

纸质出版：2026-07-15
Accepted：

丁盛,李静,杨明龙等.基于回折环引物加速LAMP与激烈火球菌Argonaute蛋白的一锅式DNA快速生物传感平台用于肉制品可视化鉴定[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2026,27(07):775-787.

Sheng DING, Jing LI, Minglong YANG, et al. DNA-based one-pot rapid biosensing platform integrating turn-back loop primer-accelerated LAMP and Pyrococcus furiosus Argonaute for meat visual authentication[J]. Journal of Zhejiang University-SCIENCE B, 2026, 27(7): 775-787. DOI: 10.1631/jzus.B2500745.
丁盛,李静,杨明龙等.基于回折环引物加速LAMP与激烈火球菌Argonaute蛋白的一锅式DNA快速生物传感平台用于肉制品可视化鉴定[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2026,27(07):775-787. DOI： 10.1631/jzus.B2500745.

Sheng DING, Jing LI, Minglong YANG, et al. DNA-based one-pot rapid biosensing platform integrating turn-back loop primer-accelerated LAMP and Pyrococcus furiosus Argonaute for meat visual authentication[J]. Journal of Zhejiang University-SCIENCE B, 2026, 27(7): 775-787. DOI: 10.1631/jzus.B2500745. DOI：

摘要

肉类掺假是全球食品安全面临的一项重大挑战，因而迫切需要开发快速的现场检测技术。在此，本研究提出了一种名为OTPA（一锅法TLAMP-PfAgo分析）的智能一锅法生物传感平台。该平台将基于折返环引物加速的环介导等温扩增技术（LAMP）（TLAMP）与激烈火球菌（

Pyrococcus furiosus

）的Argonaute蛋白（PfAgo）的序列特异性检测功能完美融合，采用一种巧妙的热激活设计，利用微晶蜡在单管内实现反应的空间隔离，从而确保了操作的无污染与流程的简化。OTPA分析法具备极高的灵敏度与特异性，在30分钟内对猪肉DNA和牛肉DNA的检测限分别低至3×10

-4

ng/μL和2×10

-4

ng/μL。此外，它成功实现了双靶标鉴定，并在市售肉制品中得到了验证，其结果与标准的PCR定性检测高度一致。值得注意的是，检测结果可在蓝光下直接通过肉眼可视化，这充分凸显了这种经济高效且简便的平台在食品安全监测的即时检测（POCT）与智能生物传感领域中所具备的巨大潜力。

Abstract

Meat adulteration is a significant global food safety challenge

creating a pressin

g need for rapid and on-site detection technologies. Herein

we present an intelligent one-pot biosensing platform termed one-pot TLAMP-PfAgo assay (OTPA) that integrates the rapid amplification of turn-back loop primer-accelerated loop-mediated isothermal amplification (LAMP) (TLAMP) with the sequence-specific detection of

Pyrococcus furiosus

Argonaute (PfAgo). This system features a clever heat-activatable design using microcrystalline wax to spatially separate reactions within a single tube

enabling contamination-free and streamlined operation. The OTPA assay achieves sensitive and specific detection

with limits of detection as low as 3×10

-4

ng/μL for pork DNA and 2×10

-4

ng/μL for beef DNA within 30 min. It successfully enables duplex target identification and has been validated with commercial meat products

showing perfect concordance with standard polymerase chain reaction (PCR)-based qualitative detection. Notably

the result can be directly visualized under blue light

underscoring the substantial potential of this cost-effective and simple platform for point-of-care testing (POCT) and intelligent biosensing in food safety surveillance.

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references

Adenuga BM , Montowska M , 2023 . A systematic review of DNA-based methods in authentication of game and less common meat species . Compr Rev Food Sci Food Saf , 22 ( 3 ): 2112 - 2160 . https://doi.org/10.1111/1541-4337.13142 https://doi.org/10.1111/1541-4337.13142

Agnoli L , Capitello R , de Salvo M , et al. , 2016 . Food fraud and consumers’ choices in the wake of the horsemeat scandal . Br Food J , 118 ( 8 ): 1898 - 1913 . https://doi.org/10.1108/BFJ-04-2016-0176 https://doi.org/10.1108/BFJ-04-2016-0176

Chaudhary V , Kajla P , Dewan A , et al. , 2022 . Spectroscopic techniques for authentication of animal origin foods . Front Nutr , 9 : 979205 . https://doi.org/10.3389/fnut.2022.979205 https://doi.org/10.3389/fnut.2022.979205

Dai YF , Wu YF , Liu GZ , et al. , 2020 . CRISPR mediated biosensing toward understanding cellular biology and point-of-care diagnosis . Angew Chem Int Ed , 59 ( 47 ): 20754 - 20766 . https://doi.org/10.1002/anie.202005398 https://doi.org/10.1002/anie.202005398

Fajardo V , González I , Rojas M , et al. , 2010 . A review of current PCR-based methodologies for the authentication of meats from game animal species . Trends Food Sci Technol , 21 ( 8 ): 408 - 421 . https://doi.org/10.1016/j.tifs.2010.06.002 https://doi.org/10.1016/j.tifs.2010.06.002

Fang X , Zhang WY , Su HH , et al. , 2025 . A LAMP-based colorimetric and fluorescence dual-channel assay for on-site identification of adulterated meat by a portable device . Food Control , 169 : 110984 . https://doi.org/10.1016/j.foodcont.2024.110984 https://doi.org/10.1016/j.foodcont.2024.110984

He HJ , da Silva Ferreira MV , Wu QY , et al. , 2024 . Portable and miniature sensors in supply chain for food authentication: a review . Crit Rev Food Sci Nutr , 65 ( 20 ): 3966 - 3986 . https://doi.org/10.1080/10408398.2024.2380837 https://doi.org/10.1080/10408398.2024.2380837

Jiang H , Li Y , Lv XF , et al. , 2023 . Recent advances in cascade isothermal amplification techniques for ultra-sensitive nucleic acid detection . Talanta , 260 : 124645 . https://doi.org/10.1016/j.talanta.2023.124645 https://doi.org/10.1016/j.talanta.2023.124645

Jiang YS , Bhadra S , Li BL , et al. , 2015 . Robust strand exchange reactions for the sequence-specific, real-time detection of nucleic acid amplicons . Anal Chem , 87 ( 6 ): 3314 - 3320 . https://doi.org/10.1021/ac504387c https://doi.org/10.1021/ac504387c

Kamruzzaman M , Makino Y , Oshita S , 2015 . Non-invasive analytical technology for the detection of contamination, adulteration, and authenticity of meat, poultry, and fish: a review . Anal Chim Acta , 853 : 19 - 29 . https://doi.org/10.1016/j.aca.2014.08.043 https://doi.org/10.1016/j.aca.2014.08.043

Li XM , Zang MW , Li D , et al. , 2023 . Meat food fraud risk in Chinese markets 2012‒2021 . npj Sci Food , 7 : 12 . https://doi.org/10.1038/s41538-023-00189-z https://doi.org/10.1038/s41538-023-00189-z

Li YR , Kou J , Han X , et al. , 2023 . Argonaute-triggered visual and rebuilding-free foodborne pathogenic bacteria detection . J Hazard Mater , 454 : 131485 . https://doi.org/10.1016/j.jhazmat.2023.131485 https://doi.org/10.1016/j.jhazmat.2023.131485

Li YR , Zhao L , Wang JL , et al. , 2024 . Argonaute-based nucleic acid detection technology: advantages, current status, challenges, and perspectives . ACS Sens , 9 ( 11 ): 5665 - 5682 . https://doi.org/10.1021/acssensors.4c01631 https://doi.org/10.1021/acssensors.4c01631

Lianou A , Papakonstantinou M , Nychas GJE , et al. , 2021 . Fraud in meat and poultry products . In: Hellberg RS, Everstine K, Sklare SA (Eds.), Food Fraud: A Global Threat with Public Health and Economic Consequences . Academic Press , Amsterdam , p. 85 - 108 . https://doi.org/10.1016/B978-0-12-817242-1.00012-9 https://doi.org/10.1016/B978-0-12-817242-1.00012-9

Liu Q , Guo X , Xun GH , et al. , 2021 . Argonaute integrated single-tube PCR system enables supersensitive detection of rare mutations . Nucleic Acids Res , 49 ( 13 ): e75 - e75 . https://doi.org/10.1093/nar/gkab274 https://doi.org/10.1093/nar/gkab274

Martineau RL , Murray SA , Ci S , et al. , 2017 . Improved performance of loop-mediated isothermal amplification assays via swarm priming . Anal Chem , 89 ( 1 ): 625 - 632 . https://doi.org/10.1021/acs.analchem.6b02578 https://doi.org/10.1021/acs.analchem.6b02578

Montowska M , Pospiech E , 2010 . Authenticity determination of meat and meat products on the protein and DNA basis . Food Rev Int , 27 ( 1 ): 84 - 100 . https://doi.org/10.1080/87559129.2010.518297 https://doi.org/10.1080/87559129.2010.518297

Muflihah , Hardianto A , Kusumaningtyas P , et al. , 2023 . DNA-based detection of pork content in food . Heliyon , 9 ( 3 ): e14418 . https://doi.org/10.1016/j.heliyon.2023.e14418 https://doi.org/10.1016/j.heliyon.2023.e14418

Müller-Maatsch J , van Ruth SM , 2021 . Handheld devices for food authentication and their applications: a review . Foods , 10 ( 12 ): 2901 . https://doi.org/10.3390/foods10122901 https://doi.org/10.3390/foods10122901

Perestam AT , Fujisaki KK , Nava O , et al. , 2017 . Comparison of real-time PCR and ELISA-based methods for the detection of beef and pork in processed meat products . Food Control , 71 : 346 - 352 . https://doi.org/10.1016/j.foodcont.2016.07.017 https://doi.org/10.1016/j.foodcont.2016.07.017

Qu C , Li YZ , Du SS , et al. , 2022 . Raman spectroscopy for rapid fingerprint analysis of meat quality and security: principles, progress and prospects . Food Res Int , 161 : 111805 . https://doi.org/10.1016/j.foodres.2022.111805 https://doi.org/10.1016/j.foodres.2022.111805

Robson K , Dean M , Brooks S , et al. , 2020 . A 20-year analysis of reported food fraud in the global beef supply chain . Food Control , 116 : 107310 . https://doi.org/10.1016/j.foodcont.2020.107310 https://doi.org/10.1016/j.foodcont.2020.107310

Sheu SC , Tsou PC , Lien YY , et al. , 2018 . Development of loop-mediated isothermal amplification (LAMP) assays for the rapid detection of allergic peanut in processed food . Food Chem , 257 : 67 - 74 . https://doi.org/10.1016/j.foodchem.2018.02.124 https://doi.org/10.1016/j.foodchem.2018.02.124

Shi JY , Ding S , Li C , et al. , 2024 . Ultrafast DNA detection based on turn-back loop primer-accelerated LAMP (TLAMP) . Anal Chim Acta , 1321 : 343041 . https://doi.org/10.1016/j.aca.2024.343041 https://doi.org/10.1016/j.aca.2024.343041

Sofos JN , 2008 . Challenges to meat safety in the 21st century . Meat Sci , 78 ( 1-2 ): 3 - 13 . https://doi.org/10.1016/j.meatsci.2007.07.027 https://doi.org/10.1016/j.meatsci.2007.07.027

Stachniuk A , Sumara A , Montowska M , et al. , 2021 . Liquid chromatography‒mass spectrometry bottom-up proteomic methods in animal species analysis of processed meat for food authentication and the detection of adulterations . Mass Spectrom Rev , 40 ( 1 ): 3 - 30 . https://doi.org/10.1002/mas.21605 https://doi.org/10.1002/mas.21605

Su FL , Zhao WT , Zhao FR , et al. , 2025 . Pyrococcus furiosus Argonaute-based fluorometric biosensor for one-tube detection of cancer-associated single nucleotide polymorphisms in microRNAs . Anal Chem , 97 ( 8 ): 4678 - 4686 . https://doi.org/10.1021/acs.analchem.4c07109 https://doi.org/10.1021/acs.analchem.4c07109

Tian T , Zhou XM , 2023 . CRISPR-based biosensing strategies: technical development and application prospects . Annu Rev Anal Chem , 16 : 311 - 332 . https://doi.org/10.1146/annurev-anchem-090822-014725 https://doi.org/10.1146/annurev-anchem-090822-014725

Vlachos A , Arvanitoyannis IS , Tserkezou P , 2016 . An updated review of meat authenticity methods and applications . Crit Rev Food Sci Nutr , 56 ( 7 ): 1061 - 1096 . https://doi.org/10.1080/10408398.2012.691573 https://doi.org/10.1080/10408398.2012.691573

Yang LX , Fu SJ , Peng XK , et al. , 2014 . Identification of pork in meat products using real-time loop-mediated isothermal amplification . Biotechnol Biotechnol Equip , 28 ( 5 ): 882 - 888 . https://doi.org/10.1080/13102818.2014.963789 https://doi.org/10.1080/13102818.2014.963789

Yang N , Zhang H , Han X , et al. , 2024 . Advancements and applications of loop-mediated isothermal amplification technology: a comprehensive overview . Front Microbiol , 15 : 1406632 . https://doi.org/10.3389/fmicb.2024.1406632 https://doi.org/10.3389/fmicb.2024.1406632

Yang ZR , Mao SY , Wang L , et al. , 2023 . CRISPR/Cas and Argonaute-based biosensors for pathogen detection . ACS Sens , 8 ( 10 ): 3623 - 3642 . https://doi.org/10.1021/acssensors.3c01232 https://doi.org/10.1021/acssensors.3c01232

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