Ultrasensitive quantification of pathogens in milliliters of beverage by filtration-based digital LAMP

Yuhua Yan; Tao Yang; Zisheng Luo; Dong Li; Li Li; Xingyu Lin

doi:10.1016/j.foodchem.2022.135226

Ultrasensitive quantification of pathogens in milliliters of beverage by filtration-based digital LAMP

Food Chem. 2023 May 15:408:135226. doi: 10.1016/j.foodchem.2022.135226. Epub 2022 Dec 16.

Authors

Yuhua Yan¹, Tao Yang¹, Zisheng Luo², Dong Li³, Li Li³, Xingyu Lin⁴

Affiliations

¹ College of Biosystems Engineering and Food Science, State Key Laboratory of Fluid Power and Mechatronic Systems, Zhejiang University, Hangzhou, China.
² College of Biosystems Engineering and Food Science, State Key Laboratory of Fluid Power and Mechatronic Systems, Zhejiang University, Hangzhou, China; Key Laboratory of Agro-Products Postharvest Handling of Ministry of Agriculture and Rural Affairs, Zhejiang University, Hangzhou, China; Ningbo Research Institute, Zhejiang University, Ningbo, China.
³ College of Biosystems Engineering and Food Science, State Key Laboratory of Fluid Power and Mechatronic Systems, Zhejiang University, Hangzhou, China; Key Laboratory of Agro-Products Postharvest Handling of Ministry of Agriculture and Rural Affairs, Zhejiang University, Hangzhou, China.
⁴ College of Biosystems Engineering and Food Science, State Key Laboratory of Fluid Power and Mechatronic Systems, Zhejiang University, Hangzhou, China; Key Laboratory of Agro-Products Postharvest Handling of Ministry of Agriculture and Rural Affairs, Zhejiang University, Hangzhou, China; Ningbo Research Institute, Zhejiang University, Ningbo, China. Electronic address: xingyu@zju.edu.cn.

PMID: 36549156
DOI: 10.1016/j.foodchem.2022.135226

Abstract

The quantitative detection of pathogens in milliliters of beverage sample requires complex preprocessing. To achieve rapid and ultrasensitive quantification of pathogens in large volume food sample, we developed a filtration-based interfacial digital LAMP (idLAMP) system, which consists of a nanoporous membrane for filtration and nanoporous hydrogel for digital amplification. Digital counting of single bacteria at the membrane surface under nanoconfinement could be achieved. The idLAMP successfully accomplished the quantitative detection of Escherichia coli in 100 mL water samples within 30 min, with wide dynamic range from 0.09 to 900 cells/mL. This technique could also be well applied to the quantification of Escherichia coli and Salmonella typhi in real beverage samples (juice, tea drinks, carbonated drinks and alcoholic drinks) without tedious sample pretreatments. With facile operation, higher specificity and sensitivity and better end-point analysis capabilities, the system has great potential in quantitative counting of single bacteria in large-volume food samples.

Keywords: Digital LAMP; Filtration; Hydrogel; Large volume; Nanoconfinement; Pathogen detection.

MeSH terms

Bacteria
Beverages
Escherichia coli*
Food
Nucleic Acid Amplification Techniques / methods
Salmonella typhi*
Sensitivity and Specificity