tetano
Editor, Senior Moderator
Biosens Bioelectron
. 2025 Sep 16:290:118002.
doi: 10.1016/j.bios.2025.118002. Online ahead of print. Imidazole-enhanced luminol/H[SUB]2[/SUB]O[SUB]2[/SUB] chemiluminescent biosensing for SARS-CoV-2 nucleocapsid protein with enzymatic regulation of hemin switch
Hang Ao[SUP] 1 [/SUP], Jinshan Xiong[SUP] 2 [/SUP], Wencheng Xiao[SUP] 3 [/SUP], Wenrui Hu[SUP] 3 [/SUP], Jie Wu[SUP] 4 [/SUP], Huangxian Ju[SUP] 5 [/SUP]
Affiliations
Owing to the continuous mutations of the coronavirus, the sensitive and accurate detection of the SARS-CoV-2 nucleocapsid protein is still necessary in clinical diagnosis. Aim to the challenge of sensitivity in chemiluminescence (CL) imaging detection and the need for high-throughput clinical screening, this work used imidazole (Im) as an enhancer for hemin-DNA/luminol/H[SUB]2[/SUB]O[SUB]2[/SUB] CL system to propose a CL imaging method for SARS-CoV-2 nucleocapsid protein detection. In this system Im could reduce the hemin-DNA catalyzed oxidation energy of luminol by H[SUB]2[/SUB]O[SUB]2[/SUB] to promote the CL emission through the accelerated breaking of O-O bond of dianionic cyclic peroxide. The enhancement mechanism was demonstrated with theoretical calculations and several characterization techniques. By combining the proximity-induced initiator release with immunological recognition, an initiator-triggered DNA nanomachine was designed to achieve the release of abundant primers for activating hemin-DNA switch. This signal amplification strategy led to strong CL emission for highly sensitive imaging detection of the target. The Im-enhanced hemin-DNA/luminol/H[SUB]2[/SUB]O[SUB]2[/SUB] CL imaging assay exhibited a linear detection range over 4 orders of magnitude with a detection limit down to 2.75 pg/mL for SARS-CoV-2 nucleocapsid protein, exhibiting promising potential of both Im sensitization and the proposed DNA nanomachine to improve the sensitivity of high-throughput CL biosensing for the early diagnosis and clinical screening of the coronavirus and other diseases.
Keywords: Chemiluminescence imaging; DNA nanomachine; DNAzyme; Immunoassay; Proximity assembly; SARS-CoV-2 nucleocapsid protein.
. 2025 Sep 16:290:118002.
doi: 10.1016/j.bios.2025.118002. Online ahead of print. Imidazole-enhanced luminol/H[SUB]2[/SUB]O[SUB]2[/SUB] chemiluminescent biosensing for SARS-CoV-2 nucleocapsid protein with enzymatic regulation of hemin switch
Hang Ao[SUP] 1 [/SUP], Jinshan Xiong[SUP] 2 [/SUP], Wencheng Xiao[SUP] 3 [/SUP], Wenrui Hu[SUP] 3 [/SUP], Jie Wu[SUP] 4 [/SUP], Huangxian Ju[SUP] 5 [/SUP]
Affiliations
- PMID: 40983044
- DOI: 10.1016/j.bios.2025.118002
Owing to the continuous mutations of the coronavirus, the sensitive and accurate detection of the SARS-CoV-2 nucleocapsid protein is still necessary in clinical diagnosis. Aim to the challenge of sensitivity in chemiluminescence (CL) imaging detection and the need for high-throughput clinical screening, this work used imidazole (Im) as an enhancer for hemin-DNA/luminol/H[SUB]2[/SUB]O[SUB]2[/SUB] CL system to propose a CL imaging method for SARS-CoV-2 nucleocapsid protein detection. In this system Im could reduce the hemin-DNA catalyzed oxidation energy of luminol by H[SUB]2[/SUB]O[SUB]2[/SUB] to promote the CL emission through the accelerated breaking of O-O bond of dianionic cyclic peroxide. The enhancement mechanism was demonstrated with theoretical calculations and several characterization techniques. By combining the proximity-induced initiator release with immunological recognition, an initiator-triggered DNA nanomachine was designed to achieve the release of abundant primers for activating hemin-DNA switch. This signal amplification strategy led to strong CL emission for highly sensitive imaging detection of the target. The Im-enhanced hemin-DNA/luminol/H[SUB]2[/SUB]O[SUB]2[/SUB] CL imaging assay exhibited a linear detection range over 4 orders of magnitude with a detection limit down to 2.75 pg/mL for SARS-CoV-2 nucleocapsid protein, exhibiting promising potential of both Im sensitization and the proposed DNA nanomachine to improve the sensitivity of high-throughput CL biosensing for the early diagnosis and clinical screening of the coronavirus and other diseases.
Keywords: Chemiluminescence imaging; DNA nanomachine; DNAzyme; Immunoassay; Proximity assembly; SARS-CoV-2 nucleocapsid protein.