Development of an aptasensor based on a fluorescent particles-modified aptamer for ochratoxin A detection.
The presented work reports a generic fluorescent aptasensing design employing carboxy-modified fluorescent particles as a signal-generating probe and magnetic particles as a solid separation support. Carboxy-modified fluorescent particles were used to modify the aptamer and act as a signal-generating probe. Magnetic beads were used as an immobilization surface to perform the function of a solid separation support. As a proof of concept, the assay was used to detect ochratoxin A (OTA). Fluorescent detection based on the displacement and competition format was performed, and the obtained results were compared. The competition-based assays were characterized with improved analytical characteristics as compared to those based on the displacement principle. The competitive fluorescent assays showed a high sensitivity where the detection limit and IC50 were 0.005 and 7.2 nM respectively. The aptasensing platform was finally demonstrated for the detection of OTA in a beer sample. However, this is a generic approach that can be very easily extended to other matrixes to determine OTA. Additionally, the proposed concept of fluorescent particles as a signal-generating probe in combination with magnetic particles can also be integrated to other fluorescent-based affinity assays.
BAE: Biocapteurs-Analyses-Environnement, Universite de Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex, 66860, France. akhtarloona@gmail.com.|Interdisciplinary Research Centre in Biomedical Materials (IRCBM), COMSATS Institute of Information Technology, Lahore, 54000, Pakistan. akhtarloona@gmail.com.
Akhtar
A
Hayat
BAE: Biocapteurs-Analyses-Environnement, Universite de Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex, 66860, France.
Rupesh K
RK
Mishra
BAE: Biocapteurs-Analyses-Environnement, Universite de Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex, 66860, France.
Gaelle
G
Catanante
BAE: Biocapteurs-Analyses-Environnement, Universite de Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex, 66860, France.
Jean Louis
JL
Marty
D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001515:Beer / Q000032:analysis* / Q000382:microbiology; D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000737:chemistry*; D057230:Limit of Detection; D059346:Magnets / Q000737:chemistry*; D009183:Mycotoxins / Q000032:analysis*; D009793:Ochratoxins / Q000032:analysis*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation
D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; D009183:Mycotoxins; D009793:Ochratoxins; C025589:ochratoxin A
Displacement/competition assays; Fluorescence detection; Magnetic particles; Modified aptamer; Ochratoxin A