Sensitivity-enhanced hydrogel digital RT-LAMP with in situ enrichment and interfacial reaction for norovirus quantification in food and water

Tao Yang; Liang Xue; Zisheng Luo; Jianhan Lin; Xinyang Zhang; Fangbin Xiao; Yuanjie Liu; Dong Li; Xingyu Lin

doi:10.1016/j.jhazmat.2025.137325

Sensitivity-enhanced hydrogel digital RT-LAMP with in situ enrichment and interfacial reaction for norovirus quantification in food and water

J Hazard Mater. 2025 Jan 21:488:137325. doi: 10.1016/j.jhazmat.2025.137325. Online ahead of print.

Authors

Tao Yang¹, Liang Xue², Zisheng Luo¹, Jianhan Lin³, Xinyang Zhang³, Fangbin Xiao¹, Yuanjie Liu³, Dong Li⁴, Xingyu Lin⁵

Affiliations

¹ College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China.
² Institute of Microbiology, Guangdong Academy of Sciences, State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Safety and Health, Key Laboratory of Big Data Technologies for Food Microbiological Safety, State Administration for Market Regulation, Guangzhou 510070, China.
³ Key Laboratory of Agricultural Information Acquisition Technology, Ministry of Agriculture and Rural Affairs, China Agricultural University, Beijing 100083, China.
⁴ College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China; The Rural Development Academy, Zhejiang University, Hangzhou 310058, China.
⁵ College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China. Electronic address: xingyu@zju.edu.cn.

PMID: 39864200
DOI: 10.1016/j.jhazmat.2025.137325

Abstract

Low levels of human norovirus (HuNoV) in food and environment present challenges for nucleic acid detection. This study reported an evaporation-enhanced hydrogel digital reverse transcription loop-mediated isothermal amplification (HD RT-LAMP) with interfacial enzymatic reaction for sensitive HuNoV quantification in food and water. By drying samples on a chamber array chip, HuNoV particles were enriched in situ. The interfacial amplification of nucleic acid at the hydrogel-chip interface was triggered after coating HD RT-LAMP system. Nanoconfined spaces in hydrogels provided a simple and rapid "digital format" to quantify single virus within 15 min. Through in situ evaporation for enrichment, the sensitivity level was increased by 20 times. The universality of the sensitivity-enhanced assay was also verified using other bacteria and virus. Furthermore, a deep learning model and smartphone app were developed for automatic amplicon analysis. Multiple actual samples, including 3 lake waters, strawberry, tap water and drinking water, were in situ enriched and detected for norovirus quantification using the chamber arrays. Therefore, the sensitivity-enhanced HD RT-LAMP is an efficient assay for testing biological hazards in food safety monitoring and environmental surveillance.

Keywords: Food and water; Hydrogel digital RT-LAMP; In situ enrichment; Interfacial enzymatic reaction; Norovirus.