Development of a heterologous enzyme-linked immunosorbent assay for organophosphorus pesticides with phage-borne peptide.
An enzyme-linked immunosorbent assay (ELISA) was developed to detect organophosphorus pesticides using a phage-borne peptide that was isolated from a cyclic 8-residue peptide phage library. The IC50values of the phage ELISA ranged from 1.4 to 92.1 μg L-1for eight organophosphorus pesticides (parathion-methyl, parathion, fenitrothion, cyanophos, EPN, paraoxon-methyl, paraoxon, fenitrooxon). The sensitivity was improved 120- and 2-fold compared to conventional homologous and heterologous ELISA, respectively. The selectivity of the phage ELISA was evaluated by measuring its cross-reactivity with 23 organophosphorus pesticides, among which eight were the main cross-reactants. The spike recoveries were between 66.1% and 101.6% for the detection of single pesticide residues of parathion-methyl, parathion and fenitrothion in Chinese cabbage, apple and greengrocery, and all of the coefficient of variation were less than or equal to 15.9%. Moreover, the phage ELISA results were validated by gas chromatography. The results indicate that isolating phage-borne peptides from phage display libraries is an alternative method for the development of a heterologous immunoassay and that the developed assay has a lower limit of detection than the chemically synthesized competitor assay.
College of Plant Protection (State & Local Joint Engineering Research Center of Green Pesticide Invention and Application), Nanjing Agricultural University, Nanjing 210095, China ; Key Laboratory of Integrated Management of Crop Diseases and Pests (Nanjing Agricultural University), Ministry of Education.
Xiude
X
Hua
College of Plant Protection (State & Local Joint Engineering Research Center of Green Pesticide Invention and Application), Nanjing Agricultural University, Nanjing 210095, China ; Key Laboratory of Integrated Management of Crop Diseases and Pests (Nanjing Agricultural University), Ministry of Education.
Xiaofeng
X
Liu
College of Plant Protection (State & Local Joint Engineering Research Center of Green Pesticide Invention and Application), Nanjing Agricultural University, Nanjing 210095, China ; Key Laboratory of Integrated Management of Crop Diseases and Pests (Nanjing Agricultural University), Ministry of Education.
Haiyan
H
Shi
Department of Entomology and UCD Cancer Center, University of California, Davis, California 95616, United States.
Yanru
Y
Wang
Department of Entomology and UCD Cancer Center, University of California, Davis, California 95616, United States.
Hee Joo
HJ
Kim
Department of Entomology and UCD Cancer Center, University of California, Davis, California 95616, United States.
Shirley J
SJ
Gee
College of Plant Protection (State & Local Joint Engineering Research Center of Green Pesticide Invention and Application), Nanjing Agricultural University, Nanjing 210095, China ; Key Laboratory of Integrated Management of Crop Diseases and Pests (Nanjing Agricultural University), Ministry of Education.
Minghua
M
Wang
Institute of Plant Protection, Jiangsu Academy of Agricultural Science, Nanjing 210014, China.
Fengquan
F
Liu
Department of Entomology and UCD Cancer Center, University of California, Davis, California 95616, United States.
Bruce D
BD
Hammock
Enzyme-linked immunosorbent assay; Multi-analyte assay; Organophosphorus pesticides; Phage-borne peptide