Development of an Indirect Competitive Chemiluminescence Enzyme-linked Immunoassay for Parathion
Polyclonal antibody(PcAb) against parathion was raised and used to develop an indirect competitive chemiluminescence enzyme-linked immunoassay(icCLEIA).The hapten was prepared from thiophosphoryl chloride after a three-step substitution reaction.The hapten was coupled to bovine serum albumin(BSA) and ovalbumin(OVA) as immounogen and coating antigen respectively by active ester method.New Zealand rabbits were immunized by the immunogen to obtain anti-parathion polyclonal antibody.Several parameters that might affect icCLEIA performance were carefully optimized.Under the optimum conditions,the linear range of the developed icCLEIA was 0.24-15.83 μg/L,the IC50 was 1.14 μg/L and the limit of detection was 0.09 μg/L.The average recovery of parathion from spiked vegetables and water samples ranged from 74.6% to 121.0%.In conclusion,the icCLEIA is a practical method for trace detection of parathion in real samples.