title: Synthesis of hapten and development of immunoassay based on monoclonal antibody for the detection of Dufulin in agricultural samples.
abstract: An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for the quantitative detection of the antiviral agent against tobacco mosaic virus (TMV). Hapten 6-[2-((methyllbenzothiazol-2-yl)-1-(2-ethoxy)-O, O-diethyl-α-aminophosphonate)acetamido)] hexanoic acid (DHS) was prepared from commercial chemicals and incorporated into the spacer arm through a carbon-carbon single bond. The prepared hapten was then coupled to carrier proteins keyhole limpet hemocyanin (KLH) to be used as an immunogen for monoclonal antibody (mAb) production together with ELISA development. This assay was further optimized by the assessment of the dependence of assay parameters on organic solvents, pH, and ionic strength. The IC50 values of the optimized assay for Dufulin and the calculated limit of detection in phosphate-buffered saline (PBS) were 9.6 ± 0.59 and 0.3 ± 0.05 ng/mL, respectively. Using the optimized assays Dufulin residues in soil and tobacco samples were determined with recovery values ranging from 81.5 to 95.3%, intra-assay variation ranging from 2.88 to 6.10%, and interassay variation ranging from 6.11 to 9.42%.
authors.affiliation: State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University , Guiyang 550025, People's Republic of China.
authors.forename: Ping
authors.initials: P
authors.lastname: Lu
authors.forename: Song
authors.initials: S
authors.lastname: Yang
authors.forename: Deyu
authors.initials: D
authors.lastname: Hu
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Ding
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Shi
mesh_terms: D000911:Antibodies, Monoclonal / Q000032:analysis; D000998:Antiviral Agents / Q000032:analysis*; D052160:Benzothiazoles / Q000032:analysis*; D004350:Drug Residues / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D006241:Haptens / Q000737:chemistry*; D010935:Plant Diseases / Q000821:virology; D010942:Plant Viruses / Q000187:drug effects / Q000502:physiology; D012989:Soil Pollutants / Q000032:analysis*; D014026:Nicotiana / Q000737:chemistry* / Q000821:virology
chemical_list: D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; D052160:Benzothiazoles; D006241:Haptens; D012989:Soil Pollutants; C576734:dufulin