数字农科院2.0

Single-stranded self-folding isothermal amplification combined with lateral flow assay for detection of influenza a (H1N1) virus

文献类型： 外文期刊

作者： Qianyu Zhou;He Sun;Mengyuan Zhou;Zebin Zhang;Jianfeng Wang;Feng Tang;Jiayu Wan

作者机构：

关键词： H1N1 virus;Isothermal amplification;Lateral flow assay;Point-of-care testing

期刊名称： Microchemical Journal

ISSN： 0026-265X

年卷期： 2026 年 221 卷

页码：

收录情况： SCIE（2025版）

摘要： The highly variable nature and significant public health threat posed by influenza A (H1N1) virus underscore the urgent need for rapid, sensitive, and point-of-care detection technologies. This study presents a novel biosensor for the direct detection of H1N1 viral RNA, integrating a self-folding hairpin (HP) probe-mediated isothermal amplification system with colloidal gold-based lateral flow strips. Unlike conventional PCR or multi-primer isothermal methods, the designed HP probe autonomously folds into a stable stem-loop structure, which, upon specific target binding, triggers a Bst DNA polymerase-mediated exponential amplification cascade without thermal cycling. This process generates biotin-labeled long-stranded DNA products that are visually detected on the test strip. Key reaction parameters—including pH, HP and biotin-dATP concentrations, Bst enzyme amount, temperature, and time—were systematically optimized. Under optimal conditions, the biosensor exhibited exceptional analytical performance: a detection limit of 2 pM, a linear range from 10 pM to 1 μM, and high specificity in discriminating H1N1 from other influenza subtypes and single-base mismatches. Compared to commercial ELISA and qPCR, this method reduced detection time to 40 min, increased sensitivity by three orders of magnitude, and extended the linear range by two orders of magnitude. Recovery rates in spiked serum samples ranged from 85.35% to 94.49%, and results from infected chicken embryo allantoic fluid aligned closely with qPCR. By combining high sensitivity, strong specificity, ease of operation, and rapid response, this biosensor provides a robust and field-deployable tool for point-of-care screening and early diagnosis of H1N1 virus.

分类号：

  • 相关文献

[1]Multiplex reverse transcription loop-mediated isothermal amplification for the simultaneous detection of CVB and CSVd in chrysanthemum. Muhammad, Imtiaz,Hong, Bo,Liu, Xing-liang,Zhao, Xi-ting,Ge, Bei-bei.

[2]Recombinase Polymerase Amplification (RPA) of CaMV-35S Promoter and nos Terminator for Rapid Detection of Genetically Modified Crops. Xu, Chao,Li, Liang,Jin, Wujun,Wan, Yusong,Li, Liang,Jin, Wujun,Wan, Yusong. 2014

[3]Current advances in the identification of plant nematode diseases: From lab assays to in-field diagnostics. Shao H.,Zhang P.,Peng D.,Huang W.,Kong L.-A.,Li C.,Liu E.,Peng H.. 2023

[4]Development of nucleic acid-based RAA test strip assay for the rapid detection of foot-and-mouth disease virus. Chen, Wenxian,Niu, Xinni,Zeng, Weijun,Fang, Yiqi,Zhu, Zixiang,Yi, Lin,Zhao, Mingqiu,Ding, Hongxing,Fan, Shuangqi,Li, Zhaoyao,Chen, Jinding. 2025

[5]A Rapid RT-RAA Assay for Visual Detection of Ebola Virus: Advancing Early Diagnosis in Resource-Limited Settings. Zhenyue Li,Jun Dai,Zitong Yang,Mingda Zhang,Xuemeng Wang,Chenchen Ge,Yi Lu,Wenhao Feng,Sihui Song,Cheng Zhang,Huan Cui,Zhendong Guo. 2025

[6]A Sensitive Aptasensor Using Biotin-Streptavidin System for Patulin Detection in Apple Juice. Xiaoqian Tang,Qi Zhang,Maria Isabel Pividori,Zhaowei Zhang,Jean Louis Marty,Gaëlle Catanante. 2022

[7]Lateral Flow Assay for the Detection of African Swine Fever Virus Antibodies Using Gold Nanoparticle-Labeled Acid-Treated p72. Wenzhuang Zhu,Kaiwen Meng,Yueping Zhang,Zhigao Bu,Dongming Zhao,Geng Meng. 2022

[8]Improvement of the sensitivity of lateral flow systems for detecting mycotoxins: Up-to-date strategies and future perspectives. Li Z.,Jallow A.,Nidiaye S.,Huang Y.,Zhang Q.,Li P.,Tang X.. 2024

[9]Detection of Vibrio parahaemolyticus by one-pot LAMP-CRISPR/Cas12b combined with immunomagnetic beads. Shiqi Wang,Yongheng Gao,Yiqin Miao,Wei Jiang,Xingyu Zhang,Yuting Zheng,Jialing Wang,Xiaoying Zhang,Hongyan Liao,Yunsheng Li,Ya Liu,Qinghai Hu. 2025

[10]H9N2 avian influenza virus diagnostics utilizing specific high-sensitivity enzymatic molecular system termed RPA-based CRISPR-Cas13a. Dalin He,Saisai Zhao,Fangfang Wang,Bingrong Wu,Feng Wei,Yubo Zhao,Xinhui Wei,Hui Ren,Meijuan Zhang,Yaru Fan,Jiahao Zhang,Shumin Yu,Yi Tang,Youxiang Diao. 2025

[11]Integration of RPA and CRISPR-Cas13a collateral activity for one-step detection of DHAV-3: A biological macromolecule-enabled diagnostic platform. Saisai Zhao,Jiahao Zhang,Shumin Yu,Dalin He,Bing Li,Yaru Fan,Guocheng Liu,Yi Tang,Youxiang Diao. 2025

[12]Rapid and visual detection of Listeria monocytogenes by combining one-pot LAMP-CRISPR/Cas12b with lateral flow assay. Liu, Xuelan,Zheng, Yuting,Chen, Zhiwei,Wang, Shiqi,Liao, Hongyan,Jia, Jianye,Wang, Guijun,Wang, Jialing,Yuan, Chunyan,Guo, Xiaoxi,Yin, Yuelan,Hu, Qinghai. 2025

[13]Integrating microneedle DNA extraction to hand-held microfluidic colorimetric LAMP chip system for meat adulteration detection. Bin Xiao,Ruiming Zhao,Nan Wang,Juan Zhang,Xiaoyun Sun,Fengchun Huang,Ailiang Chen. 2023

[14]Recent advances in the rapid detection of microRNA with lateral flow assays. Nan Wang,Juan Zhang,Bin Xiao,Xiaoyun Sun,Ruibin Xie,Ailiang Chen. 2022

[15]A Portable Electrochemical Immunosensor For H.ighly Sensitive Point-Of-Care Testing O f Genetically Modified Crops. Gao, HF, Wen, LK, Tian, J, Wu, YH, Liu, F, Lin, YJ, Hua, W, Wu, G. 2019

[16]Nuclease Triggered "Signal-On" and Amplified Fluorescent Sensing of Fumonisin B1 Incorporating Graphene Oxide and Specific Aptamer. Guo X.,Qiao Q.,Zhang M.,Fauconnier M.-L.. 2022

[17]A sensitive intelligent point-of-care test method for tert-butylhydroquinone in edible oil via a test strip with a smartphone. Dan Wang,Le Zhang,Jing Zeng,Shenling Wang,Wangwang Li,Yuanlong Sun,Wenqin Wu,Zhi Quan Tian,Zhaowei Zhang. 2024

[18]Toothpick DNA extraction combined with handheld LAMP microfluidic platform for simple and rapid meat authentication. Bin Xiao,Tianping Zhou,Nan Wang,Juan Zhang,Xiaoyun Sun,Jiaci Chen,Fengchun Huang,Junbo Wang,Nan Li,Ailiang Chen. 2024

[19]A Rapid Field-Visualization Detection Platform for Genetically Modified Soybean ‘Zhonghuang 6106’ Based on RPA-CRISPR. Ran Tao,Jihong Zhang,Lixia Meng,Jin Gao,Chaohua Miao,Weixiao Liu,Wujun Jin,Yusong Wan. 2025

[20]Nanobody-based dual-mode sensing platform for highly sensitive detection of aflatoxin B1. Qinkai Yang,Huilin Jia,Guangsheng Ge,Mengnan Wang,Shurong Li,Xuan Liu,Xueyi Fan,Kai Zheng,Xiaoqian Tang,Qi Zhang,Jianlong Wang,Ran Chen,Yanru Wang. 2025

作者其他论文 更多>>