High-Throughput Low-Background G-Quadruplex Aptamer Chemiluminescence Assay for Ochratoxin A Using a Single Photonic Crystal Microsphere.

We reported a novel hemin-G-quadruplex aptamer chemiluminescence assay platform for ochratoxin A (OTA) using the single silica photonic crystal microsphere (SPCM). The oligonucleotide A sequence containing aptamer sequences of hemin and OTA is immobilized on the surface of SPCM. The other oligonucleotide B sequence containing a partially complementary sequence with one part OTA aptamer and one part hemin aptamer is used as a blocking chain. The hybridization between chain A and chain B will be influenced by the presence or absence of OTA in the system, which will affect the bioactivity of DNAzyme. Thus, the chemiluminescence signal depends on the concentration of OTA in the samples. In the single particle assay platform, the signal/noise is remarkably enhanced, and the background signal can be ignored by separating hemin from the surface of SPCM. The limit of detection of the new method reaches to the pg/mL scale, and the linear detection range is 4 orders of magnitude for OTA. The new assay platform can provide a sensitive, cost-efficient, simple, and high-throughput screening for OTA.