Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 59, Issue 3, Pages 809-813Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jf103261u
Keywords
Ochratoxin A; ELISA; chemiluminescence; enhancement; peroxidase; soy
Funding
- Russian Foundation for Basic Research [09-03-92008-NNS_a]
- National Science Council of the Republic of China (Taiwan) [NSC-98-2923-B-040-001-MY2]
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A direct competitive chemiluminescent enzyme-linked immunosorbent assay (CL-ELISA) for the determination of ochratoxin A (OTA) was developed using soybean peroxidase (SbP) in combination with 3-(10 '-phenothiazinyl)propane-1-sulfonate (SPTZ) and 4-morpholinopyridine (MORPH) as a detection system. By varying the concentrations of the capture monoclonal anti-OTA antibody, a conjugate of OTA with SbP, and the composition of blocking buffers, the conditions of the immunoassay were optimized. Advantages of CL-ELISA were demonstrated by comparison with ELISA with colorimetric detection (COL-ELISA). The values of IC10, IC50, and working range (IC20-IC80) for CL-ELISA and COL-ELISA were 0.01, 0.08, and 0.02-0.3 ng/mL and 0.08, 0.58, and 0.17-2.2 ng/mL, respectively. The recovery values of CL-ELISA from three soybean spiked samples with OTA concentrations of 0.07, 0.1, and 0.15 ng/mL ranged from 72 to 125%. Determination of OTA in 21 various agricultural commodities showed that OTA in 8 examined samples was not detected by COL-ELISA. Furthermore, it was found that in 4 of these 8 samples the developed CL-ELISA determined OTA at levels from 0.96 to 4.64 ng/g.
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