Development of a Heterologous Enzyme-Linked Immunosorbent Assay for Organophosphorus Pesticides with Phage-Borne Peptide
HHS Public Access Author manuscript Author Manuscript Author ManuscriptRSC Adv Author Manuscript. Author manuscript; Author Manuscript available in PMC 2015 August 17. Published in final edited form as: RSC Adv. 2014 January 1; 4(80): 42445–42453. doi:10.1039/C4RA07059C. Development of a heterologous enzyme-linked immunosorbent assay for organophosphorus pesticides with phage-borne peptide Xiude Huaa,b, Xiaofeng Liua,b, Haiyan Shia,b, Yanru Wangc, Hee Joo Kimc, Shirley J. Geec, Minghua Wanga,b,*, Fengquan Liud,*, and Bruce D. Hammockc aCollege of Plant Protection (State & Local Joint Engineering Research Center of Green Pesticide Invention and Application), Nanjing Agricultural University, Nanjing 210095, China bKey Laboratory of Integrated Management of Crop Diseases and Pests (Nanjing Agricultural University), Ministry of Education cDepartment of Entomology and UCD Cancer Center, University of California, Davis, California 95616, United States dInstitute of Plant Protection, Jiangsu Academy of Agricultural Science, Nanjing 210014, China Abstract 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 −1 library. The IC50 values of the phage ELISA ranged from 1.4 to 92.1 μg L for 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%.
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