In this work, an electrochemical enzyme-linked oligonucleotide array to achieve simple and rapid multidetection of aflatoxin B1 (AFB1) is presented. The assay is based on a competitive format and disposable screen-printed cells (SPCs). Firstly, the electrodeposition of poly(aniline-anthranilic acid) copolymer (PANI-PAA) on graphite screen-printed working electrodes was performed by means of cyclic voltammetry (CV). Aflatoxin B1 conjugated with bovine serum albumin (AFB1-BSA) was then immobilized by covalent binding on PANI-PAA copolymer. After performing the affinity reaction between AFB1 and the biotinylated DNA-aptamer (apt-BIO), the solution was dropped on the modified SPCs and the competition was carried out. The biotinylated complexes formed onto the sensor surface were coupled with a streptavidin-alkaline phosphatase conjugate. 1-naphthyl phosphate was used as enzymatic substrate; the electroactive product was detected by differential pulse voltammetry (DPV). The response of the enzyme-linked oligonucleotide assay was signal-off, according to the competitive format. A dose-response curve was obtained between 0.1 ng mL−1 and 10 ng mL−1 and a limit of detection of 0.086 ng mL−1 was achieved. Finally, preliminary experiments in maize flour samples spiked with AFB1 were also performed.
Electrochemical enzyme-linked oligonucleotide array for aflatoxin B1 detection / Selvolini G.; Lettieri M.; Tassoni L.; Gastaldello S.; Grillo M.; Maran C.; Marrazza G.. - In: TALANTA. - ISSN 0039-9140. - ELETTRONICO. - 203:(2019), pp. 49-57. [10.1016/j.talanta.2019.05.044]
Electrochemical enzyme-linked oligonucleotide array for aflatoxin B1 detection
Tassoni L.;Gastaldello S.;
2019
Abstract
In this work, an electrochemical enzyme-linked oligonucleotide array to achieve simple and rapid multidetection of aflatoxin B1 (AFB1) is presented. The assay is based on a competitive format and disposable screen-printed cells (SPCs). Firstly, the electrodeposition of poly(aniline-anthranilic acid) copolymer (PANI-PAA) on graphite screen-printed working electrodes was performed by means of cyclic voltammetry (CV). Aflatoxin B1 conjugated with bovine serum albumin (AFB1-BSA) was then immobilized by covalent binding on PANI-PAA copolymer. After performing the affinity reaction between AFB1 and the biotinylated DNA-aptamer (apt-BIO), the solution was dropped on the modified SPCs and the competition was carried out. The biotinylated complexes formed onto the sensor surface were coupled with a streptavidin-alkaline phosphatase conjugate. 1-naphthyl phosphate was used as enzymatic substrate; the electroactive product was detected by differential pulse voltammetry (DPV). The response of the enzyme-linked oligonucleotide assay was signal-off, according to the competitive format. A dose-response curve was obtained between 0.1 ng mL−1 and 10 ng mL−1 and a limit of detection of 0.086 ng mL−1 was achieved. Finally, preliminary experiments in maize flour samples spiked with AFB1 were also performed.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.