An indirect competitive elisa for determination of citrinin
2011
Li, Yongning | Wang, Yuanyuan | GUO, YANGHAO
Citrinin (CIT)–protein artificial antigen was prepared by conjugating CIT with keyhole limpet hemocyanin by 1,4âbutanediol diglycidyl ether. By immunization and fusion, a hybridoma cell line named K2âF3, which stably secreted the monoclonal antibody (McAb) against CIT was obtained. The titer of the CITâ specific McAbs purified by affinity chromatography came to 1:1.38â×â105. The indirect competitive enzymeâlinked immunosorbent assay (ELISA) showed that the detection limit of CIT was 0.01âng/mL, with a good linearity ranging from 0.01 to 1.0âng/mL. The McAb was highly specific and crossâreactivity rate was less than 0.01%. The data showed that the ELISA developed could accurately determine CIT in the real contaminated red yeast rice samples. The systematic error was low with the coefficient of variation from 2.91 to 8.53% by the ELISA. PRACTICAL APPLICATIONS: CIT is a toxic fungal metabolite. It often occurs in naturally contaminated commodities, such as in corn, barley, wheat, apple and some foods. CIT is hepatonephrotoxic and implicated in disease outbreaks in animals and humans. Nowadays, food security problem due to the contamination of CIT has been a growing concern for the government and the public, especially in Asian countries. In this work, a highâspecificity McAb against citrinin was obtained. The indirect competitive ELISA based on the obtained McAb exhibited a high sensitivity and precision. The detection limit of CIT was 0.01âng/mL. The indirect competitive ELISA developed could determine CIT with a satisfactory precision for the real red yeast rice samples. This work would be helpful for establishing the technology and developing the kit to determine CITâcontaminated samples by ELISA.
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