tetano
Editor, Senior Moderator
Anal Chim Acta
. 2024 Apr 15:1298:342398.
doi: 10.1016/j.aca.2024.342398. Epub 2024 Feb 20. AIEgens-enhanced rapid sensitive immunofluorescent assay for SARS-CoV-2 with digital microfluidics
Yuping Zeng[SUP] 1 [/SUP], Xiangyu Gan[SUP] 2 [/SUP], Zhourui Xu[SUP] 3 [/SUP], Xiaoxiang Hu[SUP] 4 [/SUP], Chenxuan Hu[SUP] 5 [/SUP], Hanbin Ma[SUP] 6 [/SUP], Hangjia Tu[SUP] 7 [/SUP], Bao Chai[SUP] 8 [/SUP], Chengbin Yang[SUP] 9 [/SUP], Siyi Hu[SUP] 10 [/SUP], Yujuan Chai[SUP] 11 [/SUP]
Affiliations
Background: Sensitive and rapid antigen detection is critical for the diagnosis and treatment of infectious diseases, but conventional ELISAs including chemiluminescence-based assays are limited in sensitivity and require many operation steps. Fluorescence immunoassays are fast and convenient but often show limited sensitivity and dynamic range.
Results: To address the need, an aggregation-induced emission fluorgens (AIEgens) enhanced immunofluorescent assay with beads-based quantification on the digital microfluidic (DMF) platform was developed. Portable DMF devices and chips with small electrodes were fabricated, capable of manipulating droplets within 100 nL and boosting the reaction efficiency. AIEgen nanoparticles (NPs) with high fluorescence and photostability were synthesized to enhance the test sensitivity and detection range. The integration of AIEgen probes, transparent DMF chip design, and the large magnetic beads (10 μm) as capture agents enabled rapid and direct image-taking and signal calculation of the test result. The performance of this platform was demonstrated by point-of-care quantification of SARS-CoV-2 nucleocapsid (N) protein. Within 25 min, a limit of detection of 5.08 pg mL[SUP]-1[/SUP] and a limit of quantification of 8.91 pg mL[SUP]-1[/SUP] can be achieved using <1 μL sample. The system showed high reproducibility across the wide dynamic range (10-10[SUP]5[/SUP] pg mL[SUP]-1[/SUP]), with the coefficient of variance ranging from 2.6% to 9.8%.
Significance: This rapid, sensitive AIEgens-enhanced immunofluorescent assay on the DMF platform showed simplified reaction steps and improved performance, providing insight into the small-volume point-of-care testing of different biomarkers in research and clinical applications.
Keywords: Aggregation-induced luminescence nanoparticles; Digital microfluidics; High-sensitivity biosensor; Immunofluorescent assay; SARS-CoV-2.
. 2024 Apr 15:1298:342398.
doi: 10.1016/j.aca.2024.342398. Epub 2024 Feb 20. AIEgens-enhanced rapid sensitive immunofluorescent assay for SARS-CoV-2 with digital microfluidics
Yuping Zeng[SUP] 1 [/SUP], Xiangyu Gan[SUP] 2 [/SUP], Zhourui Xu[SUP] 3 [/SUP], Xiaoxiang Hu[SUP] 4 [/SUP], Chenxuan Hu[SUP] 5 [/SUP], Hanbin Ma[SUP] 6 [/SUP], Hangjia Tu[SUP] 7 [/SUP], Bao Chai[SUP] 8 [/SUP], Chengbin Yang[SUP] 9 [/SUP], Siyi Hu[SUP] 10 [/SUP], Yujuan Chai[SUP] 11 [/SUP]
Affiliations
- PMID: 38462346
- DOI: 10.1016/j.aca.2024.342398
Background: Sensitive and rapid antigen detection is critical for the diagnosis and treatment of infectious diseases, but conventional ELISAs including chemiluminescence-based assays are limited in sensitivity and require many operation steps. Fluorescence immunoassays are fast and convenient but often show limited sensitivity and dynamic range.
Results: To address the need, an aggregation-induced emission fluorgens (AIEgens) enhanced immunofluorescent assay with beads-based quantification on the digital microfluidic (DMF) platform was developed. Portable DMF devices and chips with small electrodes were fabricated, capable of manipulating droplets within 100 nL and boosting the reaction efficiency. AIEgen nanoparticles (NPs) with high fluorescence and photostability were synthesized to enhance the test sensitivity and detection range. The integration of AIEgen probes, transparent DMF chip design, and the large magnetic beads (10 μm) as capture agents enabled rapid and direct image-taking and signal calculation of the test result. The performance of this platform was demonstrated by point-of-care quantification of SARS-CoV-2 nucleocapsid (N) protein. Within 25 min, a limit of detection of 5.08 pg mL[SUP]-1[/SUP] and a limit of quantification of 8.91 pg mL[SUP]-1[/SUP] can be achieved using <1 μL sample. The system showed high reproducibility across the wide dynamic range (10-10[SUP]5[/SUP] pg mL[SUP]-1[/SUP]), with the coefficient of variance ranging from 2.6% to 9.8%.
Significance: This rapid, sensitive AIEgens-enhanced immunofluorescent assay on the DMF platform showed simplified reaction steps and improved performance, providing insight into the small-volume point-of-care testing of different biomarkers in research and clinical applications.
Keywords: Aggregation-induced luminescence nanoparticles; Digital microfluidics; High-sensitivity biosensor; Immunofluorescent assay; SARS-CoV-2.