Journal of Zhejiang University

Journal of Zhejiang University SCIENCE B 2026 Vol.27 No.7 P.775-787

DNA-based one-pot rapid biosensing platform integrating turn-back loop primer-accelerated LAMP and Pyrococcus furiosus Argonaute for meat visual authentication

Author(s): Sheng DING, Jing LI, Minglong YANG, Yujia SONG, Xiangke NIU, Mei LI, Yao LUO
Affiliation(s): 1. Clinical Medical College & Affiliated Hospital, Chengdu University, Chengdu 610106, China more
Corresponding email(s): luoyao@scu.edu.cn, limei9138@scu.edu.cn
Key Words: Loop-mediated isothermal amplification (LAMP), Turn-back loop primer-accelerated LAMP (TLAMP), Pyrococcus furiosus Argonaute (PfAgo), Meat authentication, One-pot assay, Point-of-care testing (POCT)

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Sheng DING, Jing LI, Minglong YANG, Yujia SONG, Xiangke NIU, Mei LI, Yao LUO. 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.

@article{title="DNA-based one-pot rapid biosensing platform integrating turn-back loop primer-accelerated LAMP and Pyrococcus furiosus Argonaute for meat visual authentication",
author="Sheng DING, Jing LI, Minglong YANG, Yujia SONG, Xiangke NIU, Mei LI, Yao LUO",
journal="Journal of Zhejiang University Science B",
volume="27",
number="7",
pages="775-787",
year="2026",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2500745"
}

%0 Journal Article
%T DNA-based one-pot rapid biosensing platform integrating turn-back loop primer-accelerated LAMP and Pyrococcus furiosus Argonaute for meat visual authentication
%A Sheng DING
%A Jing LI
%A Minglong YANG
%A Yujia SONG
%A Xiangke NIU
%A Mei LI
%A Yao LUO
%J Journal of Zhejiang University SCIENCE B
%V 27
%N 7
%P 775-787
%@ 1673-1581
%D 2026
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2500745

TY - JOUR
T1 - DNA-based one-pot rapid biosensing platform integrating turn-back loop primer-accelerated LAMP and Pyrococcus furiosus Argonaute for meat visual authentication
A1 - Sheng DING
A1 - Jing LI
A1 - Minglong YANG
A1 - Yujia SONG
A1 - Xiangke NIU
A1 - Mei LI
A1 - Yao LUO
J0 - Journal of Zhejiang University Science B
VL - 27
IS - 7
SP - 775
EP - 787
%@ 1673-1581
Y1 - 2026
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2500745

Abstract
Chinese Summary
Academic Network
Reviewer Comment

Abstract: Meat adulteration is a significant global food safety challenge, creating a pressing 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.

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¹，李静¹，杨明龙¹，宋雨佳¹，牛翔科¹，李美²，罗耀²
¹成都大学临床医学院附属医院，中国成都，610106
²四川大学华西医院四川医学检验临床医学研究中心检验医学系，中国成都，610041
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。

关键词：环介导等温扩增（LAMP）；回折环引物加速LAMP（TLAMP）；激烈火球菌Argonaute蛋白（PfAgo）；肉类鉴定；一管化检测；现场检测（POCT）

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]AdenugaBM, MontowskaM, 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.

[2]AgnoliL, CapitelloR, de SalvoM, et al., 2016. Food fraud and consumers’ choices in the wake of the horsemeat scandal. Br Food J, 118(8):1898-1913.

[3]ChaudharyV, KajlaP, DewanA, et al., 2022. Spectroscopic techniques for authentication of animal origin foods. Front Nutr, 9:979205.

[4]DaiYF, WuYF, LiuGZ, et al., 2020. CRISPR mediated biosensing toward understanding cellular biology and point-of-care diagnosis. Angew Chem Int Ed, 59(47):20754-20766.

[5]FajardoV, GonzálezI, RojasM, 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.

[6]FangX, ZhangWY, SuHH, 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.

[7]HeHJ, da Silva FerreiraMV, WuQY, et al., 2024. Portable and miniature sensors in supply chain for food authentication: a review. Crit Rev Food Sci Nutr, 65(20):3966-3986.

[8]JiangH, LiY, LvXF, et al., 2023. Recent advances in cascade isothermal amplification techniques for ultra-sensitive nucleic acid detection. Talanta, 260:124645.

[9]JiangYS, BhadraS, LiBL, et al., 2015. Robust strand exchange reactions for the sequence-specific, real-time detection of nucleic acid amplicons. Anal Chem, 87(6):3314-3320.

[10]KamruzzamanM, MakinoY, OshitaS, 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.

[11]LiXM, ZangMW, LiD, et al., 2023. Meat food fraud risk in Chinese markets 2012‒2021. npj Sci Food, 7:12.

[12]LiYR, KouJ, HanX, et al., 2023. Argonaute-triggered visual and rebuilding-free foodborne pathogenic bacteria detection. J Hazard Mater, 454:131485.

[13]LiYR, ZhaoL, WangJL, et al., 2024. Argonaute-based nucleic acid detection technology: advantages, current status, challenges, and perspectives. ACS Sens, 9(11):5665-5682.

[14]LianouA, PapakonstantinouM, NychasGJE, 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.

[15]LiuQ, GuoX, XunGH, et al., 2021. Argonaute integrated single-tube PCR system enables supersensitive detection of rare mutations. Nucleic Acids Res, 49(13):e75-e75.

[16]MartineauRL, MurraySA, CiS, et al., 2017. Improved performance of loop-mediated isothermal amplification assays via swarm priming. Anal Chem, 89(1):625-632.

[17]MontowskaM, PospiechE, 2010. Authenticity determination of meat and meat products on the protein and DNA basis. Food Rev Int, 27(1):84-100.

[18]Muflihah, HardiantoA, KusumaningtyasP, et al., 2023. DNA-based detection of pork content in food. Heliyon, 9(3):e14418.

[19]Müller-MaatschJ, van RuthSM, 2021. Handheld devices for food authentication and their applications: a review. Foods, 10(12):2901.

[20]PerestamAT, FujisakiKK, NavaO, 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.

[21]QuC, LiYZ, DuSS, et al., 2022. Raman spectroscopy for rapid fingerprint analysis of meat quality and security: principles, progress and prospects. Food Res Int, 161:111805.

[22]RobsonK, DeanM, BrooksS, et al., 2020. A 20-year analysis of reported food fraud in the global beef supply chain. Food Control, 116:107310.

[23]SheuSC, TsouPC, LienYY, 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.

[24]ShiJY, DingS, LiC, et al., 2024. Ultrafast DNA detection based on turn-back loop primer-accelerated LAMP (TLAMP). Anal Chim Acta, 1321:343041.

[25]SofosJN, 2008. Challenges to meat safety in the 21st century. Meat Sci, 78(1-2):3-13.

[26]StachniukA, SumaraA, MontowskaM, 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.

[27]SuFL, ZhaoWT, ZhaoFR, 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.

[28]TianT, ZhouXM, 2023. CRISPR-based biosensing strategies: technical development and application prospects. Annu Rev Anal Chem, 16:311-332.

[29]VlachosA, ArvanitoyannisIS, TserkezouP, 2016. An updated review of meat authenticity methods and applications. Crit Rev Food Sci Nutr, 56(7):1061-1096.

[30]YangLX, FuSJ, PengXK, et al., 2014. Identification of pork in meat products using real-time loop-mediated isothermal amplification. Biotechnol Biotechnol Equip, 28(5):882-888.

[31]YangN, ZhangH, HanX, et al., 2024. Advancements and applications of loop-mediated isothermal amplification technology: a comprehensive overview. Front Microbiol, 15:1406632.

[32]YangZR, MaoSY, WangL, et al., 2023. CRISPR/Cas and Argonaute-based biosensors for pathogen detection. ACS Sens, 8(10):3623-3642.

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