title: Development of an enzyme-linked immunosorbent assay for detection of clopidol residues in chicken tissues.
abstract: BACKGROUND
  Clopidol is mainly used for the prevention and treatment of coccidiosis, which poses a serious potential hazard to public health, in veterinary medicine. The aim of this study was to prepare monoclonal antibodies (mAbs) against clopidol (CLOP) and develop an immunoassay for detecting CLOP residues in chicken tissues. After derivation, CLOP hapten was conjugated to carrier proteins to synthesize the artificial antigen, and immunized Balb/C mice were employed to screen mAbs.
  
  
  RESULTS
  A sensitive hybridoma named C1G3 was screened out and two indirect competitive enzyme-linked immunosorbent assay (icELISA) standard curves were established. For the traditional two-step assay the linear range was from 0.06 to 98 ng mL(-1) , with half-maximal inhibitory concentration (IC50 ) and limit of detection (LOD) values of 2.76 ng mL(-1) and 0.03 ng mL(-1) respectively, while the rapid one-step icELISA had a working range from 0.08 to 102 ng mL(-1) , with IC50 and LOD values of 3.52 ng mL(-1) and 0.03 ng mL(-1) respectively. It was also indicated that a 10-fold dilution in chicken muscles gave an inhibition curve almost the same as that obtained in phosphate-buffered saline. When applied to spiking tests in chicken samples, the correlation coefficient (R(2) ) between concentrations added and measured was 0.9534.
  
  
  CONCLUSIONS
  The results of this study suggest that the immunoassay described is a promising alternative for screening CLOP residues in biological matrices and is suitable for routine diagnostics.
authors.affiliation: Henan Institute of Science and Technology, Xinxiang, 453003, China.
authors.forename: Jin-Qing
authors.initials: JQ
authors.lastname: Jiang
authors.forename: Hai-Tang
authors.initials: HT
authors.lastname: Zhang
authors.forename: Hui-Hui
authors.initials: HH
authors.lastname: Zhang
authors.forename: Zi-Liang
authors.initials: ZL
authors.lastname: Wang
authors.forename: Xue-Feng
authors.initials: XF
authors.lastname: Yang
authors.forename: Guo-Ying
authors.initials: GY
authors.lastname: Fan
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002645:Chickens; D003007:Clopidol / Q000032:analysis*; D003049:Coccidiostats / Q000032:analysis; D004032:Diet; D004350:Drug Residues / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards*; D005260:Female; D005506:Food Contamination / Q000032:analysis*; D006241:Haptens / Q000276:immunology; D006825:Hybridomas; D008460:Meat / Q000032:analysis*; D008807:Mice, Inbred BALB C; D009132:Muscles / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D003049:Coccidiostats; D006241:Haptens; D003007:Clopidol
keywords: artificial antigen; chicken tissues; clopidol; immunoassay; monoclonal antibody