Role of background observed in aptasensor with chemiluminescence detection

One-step chemiluminescent aptasensor was developed using chemically initiated electron exchange luminescence (CIEEL) between high-energy intermediate formed from 1,1′-oxalyldiimidazole chemiluminescence (ODI-CL) reaction and G-quadruplex (ochratoxin A (OTA)-bound aptamer conjugated with TEX615) generated. The sensitivity of chemiluminescent aptasensor, optimized with various variables (e.g., property of microfibers fabricated with 3,4,9,10-perylenetetracarboxylic dimide, determination of fluorescent dye labeled with aptamer, physical properties of buffer solution), was dependent on the background (concentration of high-energy intermediate) generated in ODI-CL reaction. The limit of detection (LOD=background+3×standard deviation, 0.5 nM) of ODI-CL aptasensor with lower background was lower than that (3.7 nM) with 20 times higher background. Also, the ratio of signal to background (S/B) of ODI-CL aptasensor with low background was about 5-fold higher than that with high background. The sensitivities of ODI-CL aptasensors, with low as well as high background, capable of accurately and precisely quantifying OTA within 10 min, were better than those of fluorescent aptasensors and as good as those of highly sensitive but time-consuming competitive enzyme-linked immune-sorbent assays (ELISAs) using expensive antibody produced with the sacrifice of small animals.