Multiplex chemiluminescent immunoassay for screening of mycotoxins using photonic crystal microsphere suspension array

Abstract

A novel multiplex chemiluminescent mycotoxin immunoassay suspension array system was developed by combining the silica photonic crystal microspheres (SPCMs) encoding technique and a chemiluminescent immunoassay (CLIA) method. The SPCMs were used as a carrier of the suspension array and encoded by their reflectance peak positions, which overcome fluorescence photobleaching, and the potential interference between the encoding fluorescence and detection fluorescence. Aflatoxin B1 (AFB1), fumonisin B1 (FB1) and ochratoxin A (OTA) artificial antigens were immobilized on the surfaces of SPCMs by using 3-glycidoxypropyltrimethoxysilane as a linker. Horseradish peroxidase (HRP) was used as a labeling enzyme for the secondary antibody in the enzyme-catalyze H2O2-luminol chemiluminescence system. The CLIA detection system was easily integrated with a multifunctional microplate reader and displayed a two to three orders of magnitude dynamic linear detection range from 0.001 to 1, 0.001 to 1, and 0.01 to 1 ng mL(-1) for AFB1, FB1 and OTA with 50% inhibitory concentrations (IC50) of 0.01, 0.036, and 0.04 ng mL(-1), respectively. The recovery rates are in the range of 63.5 to 121.6% for the three mycotoxins in three kinds of spiked cereal samples. The results of detection in 12 naturally contaminated cereal samples were consistent with that of the classic enzyme-linked immunosorbent assay (ELISA) method. This proposed system is simple, rapid, low cost and high throughput for multiplex mycotoxin assay.

Green preparation of chlorine-doped graphene and its application in electrochemical sensor for chloramphenicol detection

“…Nowadays, various conventional analytical methods, such as liquid chromatography-mass spectroscopy (LC-MS) [4], gas chromatography-mass spectroscopy (GC-MS) [5], capillary electrophoresis (CE) [6], and chemiluminescent immunoassay [7], have been put forward for CAP residue detection. Nevertheless, complicated pretreatment processes, expensive instruments and professional operator requirements make these methods not suitable to routine and rapid analysis of samples.…”

Mycotoxins Contaminant in Kelp: A Neglected Dietary Exposure Pathway

“…Antibody-based immunoassays were also applied for mycotoxin detection with advantages of simplicity, low–cost and high throughput. These immunoassays mainly include enzyme linked immunosorbent assay (ELISA) [ 23 , 24 , 25 ], fluorescence polarization immunoassay (FPIA) [ 26 , 27 , 28 ], surface plasmon resonance (SPR) [ 29 , 30 , 31 , 32 ], flow cytometric microsphere immunoassay [ 33 , 34 , 35 ], and rapid strip tests [ 36 , 37 ]. However, these methods mainly focused on cereal matrix and related products.…”

High Throughput Detection Methods for Multiplex Mycotoxins

“…For example, the fluorescence dyes tend to be quenched or bleached and the optic system are complicated [79,81]. Our group established the easier, simpler and more flexible suspension array system which is based on silica photonic crystal microsphere (SPCM) encoding with their structure colors for multiplex mycotoxin analysis [79][80][81][82].…”

“…Our group established the easier, simpler and more flexible suspension array system which is based on silica photonic crystal microsphere (SPCM) encoding with their structure colors for multiplex mycotoxin analysis [79][80][81][82]. SPCMs for multiplex mycotoxin analysis showed a wide detection linear range and pg/mL LOD [79,81].…”