Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 398, Issue 5, Pages 2125-2132Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-010-4146-1
Keywords
Electrochemiluminescence; Aptamer; Ochratoxin A; Gold-nanoparticle-modified gold electrode
Funding
- 863 project [2008AA10Z419]
- NSFC [20805019]
- Ministry of Education [20070295014]
- NSF of Jiangsu Province [BK20081603]
- 111 project [B07029]
- Scientific and Technical Development Project of Qingdao in China [09-1-3-45-jch]
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A highly selective electrochemiluminescent biosensor for the detection of target nephrotoxic toxin, ochratoxin A (OTA), was developed using a DNA aptamer as the recognition element and N-(4-aminobutyl)-N-ethylisoluminol (ABEI) as the signal-producing compound. The electrochemiluminescent aptamer biosensor was fabricated by immobilizing aptamer complementary DNA 1 sequence onto the surface of a gold-nanoparticle (AuNP)-modified gold electrode. ABEI-labeled aptamer DNA 2 sequence hybridized to DNA 1 and was utilized as an electrochemiluminescent probe. A decreased electrochemiluminescence (ECL) signal was generated upon aptamer recognition of the target OTA, which induced the dissociation of DNA 2 (ABEI-labeled aptamer electrochemiluminescent probe) from DNA 1 and moved it far away from the electrode surface. Under the optimal conditions, the decreased ECL intensity was proportional to an OTA concentration ranging from 0.02 to 3.0 ng mL(-1), with a detection limit of 0.007 ng mL(-1). The relative standard deviation was 3.8% at 0.2 ng mL(-1) (n = 7). The proposed method has been applied to measure OTA in naturally contaminated wheat samples and validated by an official method. This work demonstrates the combination of a highly binding aptamer with a highly sensitive ECL technique to design an electrochemiluminescent biosensor, which is a very promising approach for the determination of small-molecule toxins. <
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