Nanopore-assisted ELISA for ultrasensitive, portable, and on-site detection of ricin

Jianing Chen; Zhuoqun Su; Wenrui Li; Ziye Pei; Di Wu; Lin Li; Yongning Wu; Guoliang Li

doi:10.1016/j.talanta.2024.127136

Nanopore-assisted ELISA for ultrasensitive, portable, and on-site detection of ricin

Talanta. 2024 Oct 31:283:127136. doi: 10.1016/j.talanta.2024.127136. Online ahead of print.

Authors

Jianing Chen¹, Zhuoqun Su², Wenrui Li¹, Ziye Pei¹, Di Wu³, Lin Li⁴, Yongning Wu⁵, Guoliang Li⁶

Affiliations

¹ School of Food Science and Engineering, Shaanxi University of Science and Technology, Xi'an, 710021, China.
² School of Food Science and Engineering, Shaanxi University of Science and Technology, Xi'an, 710021, China. Electronic address: zhuoqunsu@sust.edu.cn.
³ Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, 19 Chlorine Gardens, Belfast BT9 5DL, United Kingdom.
⁴ Animal-Derived Food Safety Innovation Team, College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China.
⁵ NHC Key Laboratory of Food Safety Risk Assessment, China National Center for Food Safety Risk Assessment, Beijing, 100017, China.
⁶ School of Food Science and Engineering, Shaanxi University of Science and Technology, Xi'an, 710021, China. Electronic address: 61254368@163.com.

PMID: 39532053
DOI: 10.1016/j.talanta.2024.127136

Abstract

Effective detection technologies in food safety with the merits of portable and on-site detection potential are always in pressing demand. Herein, we developed a nanopore-assisted Enzyme-linked immunosorbent assay (NELISA) platform, which innovatively introduced hairpin DNA (HP DNA) probes as reaction substrates. This innovation of substrates effectively avoided the inherent limitations of colorimetric signals (i.e., low sensitivity and inaccurate results) and greatly improved the sensitivity and accuracy of NELISA platform. The alkaline phosphatase (ALP)-modified detection antibody (ALP-Ab2) can specifically bind to ricin and hydrolyze the phosphate groups modified on the HP DNA probes. Nanopore recordings demonstrated that two states of probes produced highly distinguishable nanopore events, enabling the qualitative and quantitative detection of ricin. This NELISA platform fully combined the specificity of ELISA with the ultra-sensitivity, and unique single-molecule fingerprint recognition of nanopore, showing a great on-site detection potential. This method achieved the ultrasensitive and reliable detection of ricin down to 2.46 fg/mL, which enhanced the detection sensitivity by at least 10⁶-fold compared to traditional ELISA. Furthermore, the proposed method was capable of accurately detecting ricin in real food samples with satisfactory recoveries.

Keywords: ELISA; Food safety; NELISA; Nanopore; Ricin.