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

• Published in: Talanta (Oxford) Vol. 283; p. 127136
• Main Authors: Chen, Jianing, Su, Zhuoqun, Li, Wenrui, Pei, Ziye, Wu, Di, Li, Lin, Wu, Yongning, Li, Guoliang
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01-02-2025
• Subjects: ELISA; Food safety; Nanopore; NELISA; Ricin; Food safety; Nanopore; NELISA; Ricin; ELISA
• ISSN: 0039-9140; 1873-3573; 1873-3573
• DOI: 10.1016/j.talanta.2024.127136

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.
•Developed a novel nanopore-assisted ELISA (NELISA) platform combining the specificity of ELISA with the ultra-sensitivity and single-molecule recognition capability of nanopore technology.•Achieved ultrasensitive detection of ricin down to 2.46 fg/mL, demonstrating a sensitivity enhancement of at least 106-fold compared to traditional ELISA.•Successfully validated the NELISA platform for rapid, portable, and on-site detection of ricin in real food samples, overcoming limitations of traditional colorimetric ELISA methods.