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Sensitive Quantification of Aflatoxin B1 in Animal Feeds, Corn Feed Grain, and Yellow Corn Meal Using Immunomagnetic Bead-Based Recovery and Real-Time Immunoquantitative-PCR

Department of Animal Science and the Robert M. Kerr Food & Agricultural Products Center, 109 FAPC Building, Monroe Street, Oklahoma State University, Stillwater, OK 74078-6055, USA
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Present Address: Department of Toxicology, School of Pharmacy, College of Health and Pharmaceutical Sciences, University of Louisiana at Monroe, Monroe, LA 71209-0497, USA
Toxins 2014, 6(12), 3223-3237; https://doi.org/10.3390/toxins6123223
Received: 15 September 2014 / Revised: 3 November 2014 / Accepted: 25 November 2014 / Published: 2 December 2014
Aflatoxins are considered unavoidable natural mycotoxins encountered in foods, animal feeds, and feed grains. In this study, we demonstrate the application of our recently developed real-time immunoquantitative PCR (RT iq-PCR) assay for sensitive detection and quantification of aflatoxins in poultry feed, two types of dairy feed (1 and 2), horse feed, whole kernel corn feed grains, and retail yellow ground corn meal. Upon testing methanol/water (60:40) extractions of the above samples using competitive direct enzyme linked immunosorbent assay, the aflatoxin content was found to be <20 μg/kg. The RT iq-PCR assay exhibited high antigen hook effect in samples containing aflatoxin levels higher than the quantification limits (0.1–10 μg/kg), addressed by comparing the quantification results of undiluted and diluted extracts. In testing the reliability of the immuno-PCR assay, samples were spiked with 200 μg/kg of aflatoxin B1, but the recovery of spiked aflatoxin was found to be poor. Considering the significance of determining trace levels of aflatoxins and their serious implications for animal and human health, the RT iq-PCR method described in this study can be useful for quantifying low natural aflatoxin levels in complex matrices of food or animal feed samples without the requirement of extra sample cleanup. View Full-Text
Keywords: aflatoxin B1; immunomagnetic bead capture; real-time immunoquantitative PCR; animal feeds; feed grains aflatoxin B1; immunomagnetic bead capture; real-time immunoquantitative PCR; animal feeds; feed grains
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Babu, D.; Muriana, P.M. Sensitive Quantification of Aflatoxin B1 in Animal Feeds, Corn Feed Grain, and Yellow Corn Meal Using Immunomagnetic Bead-Based Recovery and Real-Time Immunoquantitative-PCR. Toxins 2014, 6, 3223-3237.

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