Chemiluminescence Immunoassay For Chloramphenicol

Here, we describe a simple, solid-phase chemiluminescence immunoassay (CLIA) for the measurement of Chloramphenicol(CAP) in foodstuffs. A rabbit anti-CAP IgG is passively adsorbed onto the walls of polypropylene plates. The labeled conjugant is horseradish peroxidase (HRP) conjugate of CAP. Luminol solution is used as the substrate of HRP. The light yield is inversely proportional to the concentration of CAP.The CLIA substrate is one of the most important prescription in CLIA. The CLIA substrate must luminesce stably and sensitively, with moderate signal, low background. We compared the self-prescription with the Roche CLIA substrate, then we found that each has its virtue with the two kinds of substrate.The HRP conjugate of CAP was diluted to 1:10000. Every well was added labeled CAP(50 μ I) and standards or samples(100 μ I), then plates were placed in 37℃ for 1h. After washing, each well was added 180 μ I CLIA substrate. The result was obtained by luminometer after adding substrate for 3 mins.The method has a similar sensitivity (0.046ng/ml), specificity, precision, and accuracy to a conventional enzyme immunoassay (EIA). The intra-assay and inter-assay CVs of ten samples were <8% and <20%, respectively, and the analytical recovery of the method was 87.4-101%. The experimental correlation coefficient of dilution was found to be 0.999 using milk supernatant as buffer. The assay range for the method was 0.05-10ng/ml, and it displayed good linearity. Compared with EIA kits of R-Biopharm, the correlative equation is y=0.996x+0.144, and r=0.987. The method overcomes the obstacles which the impurity from foodstuff can interfere the luminescent, so that we can make it possible to apply CLIA in the field of foodstuff. This method for measuring CAP is sensitive, convenient and rapid.