title: Comparison of enzyme-linked immunosorbent assays with chemiluminescent and colorimetric detection for the determination of ochratoxin A in food.
abstract: 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.
authors.affiliation: Department of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan.
authors.forename: Feng-Yih
authors.initials: FY
authors.lastname: Yu
authors.forename: Marina M
authors.initials: MM
authors.lastname: Vdovenko
authors.forename: Jing-Jhih
authors.initials: JJ
authors.lastname: Wang
authors.forename: Ivan Yu
authors.initials: IY
authors.lastname: Sakharov
mesh_terms: D002273:Carcinogens / Q000032:analysis*; D003124:Colorimetry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005504:Food Analysis / Q000379:methods*; D008163:Luminescent Measurements / Q000379:methods*; D009793:Ochratoxins / Q000032:analysis*; D012680:Sensitivity and Specificity
chemical_list: D002273:Carcinogens; D009793:Ochratoxins; C025589:ochratoxin A