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Toxins 2016, 8(1), 5; doi:10.3390/toxins8010005

Development of an Enzyme-Linked Immunosorbent Assay Method Specific for the Detection of G-Group Aflatoxins

1,2,3,4,†
,
1,5,†
,
1,†
,
1,2,3,4
,
1,2,3,4
,
1,2,3,4
,
1,2,3,4
,
6
and
1,2,3,4,*
1
Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China
2
Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China
3
Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, China
4
Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan 430062, China
5
College of Food Science and Technology, Agricultural University of Hebei, Baoding 071001, China
6
Southern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, New Orleans, LA 70124, USA
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editors: Michelangelo Pascale and Maria C. DeRosa
Received: 31 August 2015 / Revised: 9 December 2015 / Accepted: 11 December 2015 / Published: 28 December 2015
(This article belongs to the Collection Biorecognition Assays for Mycotoxins)
View Full-Text   |   Download PDF [1484 KB, uploaded 28 December 2015]   |  

Abstract

To detect and monitor G-group aflatoxins in agricultural products, we generated class-specific monoclonal antibodies that specifically recognized aflatoxins G1 and G2. Of the final three positive and stable hybridomas obtained, clone 2G6 produced a monoclonal antibody that had equal sensitivity to aflatoxins G1 and G2, and did not cross-react with aflatoxins B1, B2, or M1. Its IC50 values for aflatoxins G1 and G2 were 17.18 ng·mL−1 and 19.75 ng·mL−1, respectively. Using this new monoclonal antibody, we developed a competitive indirect enzyme-linked immunosorbent assay (CI-ELISA); the method had a limit of detection of 0.06 ng·mL−1. To validate this CI-ELISA, we spiked uncontaminated peanut samples with various amounts of aflatoxins G1 and G2 and compared recovery rates with those determined by a standard HPLC method. The recovery rates of the CI-ELISA ranging from 94% to 103% were comparable to those of the HPLC (92% to 102%). We also used both methods to determine the amounts of G-group aflatoxins in five peanut samples contaminated by aflatoxin B1-positive, and their relative standard deviations ranged from 8.4% to 17.7% (under 20%), which demonstrates a good correlation between the two methods. We further used this CI-ELISA to assess the ability of 126 fungal strains isolated from peanuts or field soils to produce G-group aflatoxins. Among these, seven stains producing different amounts of G-group aflatoxins were identified. Our results showed that the monoclonal antibody 2 G6-based CI-ELISA was suitable for the detection of G-group aflatoxins present in peanuts and also those produced by fungi. View Full-Text
Keywords: aflatoxin G1; class-specific; monoclonal antibody; ELISA; Aspergillus flavus aflatoxin G1; class-specific; monoclonal antibody; ELISA; Aspergillus flavus
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Li, P.; Zhou, Q.; Wang, T.; Zhou, H.; Zhang, W.; Ding, X.; Zhang, Z.; Chang, P.-K.; Zhang, Q. Development of an Enzyme-Linked Immunosorbent Assay Method Specific for the Detection of G-Group Aflatoxins. Toxins 2016, 8, 5.

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