Establishment of a Chemiluminescence Immunoassay Combined with Immunomagnetic Beads for Rapid Analysis Of Ochratoxin A

J AOAC Int. 2021 Oct 2:qsab104. doi: 10.1093/jaoacint/qsab104. Online ahead of print.


BACKGROUND: Ochratoxins A (OTA), as the harmful mycotoxins, are toxic secondary metabolite produced by several species of Aspergillus and Penicillium. Hence, it is of vital important to establish a sensitive method to detect OTA in various grains.

OBJECTIVE: Herein a novel highly sensitive chemiluminescence immunoassay was developed to determination of ochratoxin A in wheat, corn, and poultry feed.

METHODS: The immunomagnetic beads were used as solid phase carrier and separator in the competitive chemiluminescence method, the monoclonal antibodies anti-OTA coated on immunomagnetic beads were applied as the capturing antibody, in which OTA would compete with Ochratoxin A-alkaline phosphatase (OTA-ALP) in binding with OTA antibodies.

RESULTS: The proposed method exhibited good linearity (R2 ≥ 0.999), adequate OTA recovery rate (83.60% to 102.50%), and good repeatability. The sensitivity of the proposed method was 2.05 pg/mL. In addition, the method was highly selective, it did not have cross-reaction with other mycotoxin (e.g., AFB1, DON, ZEN). Furthermore, the developed method was applied in the analysis of wheat, corn and two animal feeds successfully, the results obtained by the chemiluminescence method showed the high correlation with those obtained by HPLC method (correlation coefficient 0.9958) when testing each extract by both methods.

CONCLUSIONS: This strategy shows great potential application for other toxic and harmful mycotoxin detection.

HIGHLIGHTS: A novel highly rapid and sensitive chemiluminescence immunoassay was established to quantitation for ochratoxin A.

PMID:34599815 | DOI:10.1093/jaoacint/qsab104

Source: Industry