title: Development of an Enzyme-Linked Immunosorbent Assay Method Specific for the Detection of G-Group Aflatoxins.
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.
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China. peiwuli@oilcrops.cn.|Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China. peiwuli@oilcrops.cn.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, China. peiwuli@oilcrops.cn.|Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan 430062, China. peiwuli@oilcrops.cn.
authors.forename: Peiwu
authors.initials: P
authors.lastname: Li
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China. zhouqianyz513@126.com.|College of Food Science and Technology, Agricultural University of Hebei, Baoding 071001, China. zhouqianyz513@126.com.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China. wangting963@163.com.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China. zhouhaiyan@caas.cn.|Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China. zhouhaiyan@caas.cn.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, China. zhouhaiyan@caas.cn.|Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan 430062, China. zhouhaiyan@caas.cn.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Zhou
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China. zhangwen@oilcrops.cn.|Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China. zhangwen@oilcrops.cn.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, China. zhangwen@oilcrops.cn.|Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan 430062, China. zhangwen@oilcrops.cn.
authors.forename: Wen
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China. dingxiaoxia@caas.cn.|Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China. dingxiaoxia@caas.cn.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, China. dingxiaoxia@caas.cn.|Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan 430062, China. dingxiaoxia@caas.cn.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Ding
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China. zhangzhaowei@caas.cn.|Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China. zhangzhaowei@caas.cn.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, China. zhangzhaowei@caas.cn.|Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan 430062, China. zhangzhaowei@caas.cn.
authors.forename: Zhaowei
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Southern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, New Orleans, LA 70124, USA. perngkuang.chang@ars.usda.gov.
authors.forename: Perng-Kuang
authors.initials: PK
authors.lastname: Chang
authors.affiliation: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China. zhangqi01@caas.cn.|Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China. zhangqi01@caas.cn.|Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, China. zhangqi01@caas.cn.|Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan 430062, China. zhangqi01@caas.cn.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000348:Aflatoxins / Q000032:analysis* / Q000276:immunology; D000818:Animals; D000906:Antibodies / Q000097:blood / Q000276:immunology; D010367:Arachis / Q000382:microbiology; D001230:Aspergillus / Q000302:isolation & purification; D045744:Cell Line, Tumor; D015169:Colony Count, Microbial; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D008807:Mice, Inbred BALB C; D009754:Nuts / Q000737:chemistry / Q000382:microbiology; D010047:Ovalbumin / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000276:immunology
chemical_list: D000348:Aflatoxins; D000906:Antibodies; C027955:aflatoxin G1; D012710:Serum Albumin, Bovine; C029754:aflatoxin G2; D010047:Ovalbumin
keywords: Aspergillus flavus; ELISA; aflatoxin G1; class-specific; monoclonal antibody