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A magnetic particles-based chemiluminescence enzyme immunoassay for rapid detection of ovalbumin
Authors:Xiao-Li Feng  Hong-Lin Ren  Yan-Song Li  Pan Hu  Yu Zhou  Zeng-Shan Liu  Dong-Ming Yan  Qi Hui  Dong Liu  Chao Lin  Nan-Nan Liu  Yan-Yan Liu  Shi-Ying Lu
Affiliation:1. Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, People’s Republic of China;2. Wuhu Kang Wei Biological Technology, Wuhu 241000, People’s Republic of China;3. Changchun Institute of Biological Products, Changchun 130062, People’s Republic of China
Abstract:Egg allergy is an important public health and safety concern, so quantification and administration of food or vaccines containing ovalbumin (OVA) are urgently needed. This study aimed to establish a rapid and sensitive magnetic particles–chemiluminescence enzyme immunoassay (MPs–CLEIA) for the determination of OVA. The proposed method was developed on the basis of a double antibodies sandwich immunoreaction and luminol–H2O2 chemiluminescence system. The MPs served as both the solid phase and separator, the anti-OVA MPs-coated polyclonal antibodies (pAbs) were used as capturing antibody, and the horseradish peroxidase (HRP)-labeled monoclonal antibody (mAb) was taken as detecting antibody. The parameters of the method were evaluated and optimized. The established MPs–CLEIA method had a linear range from 0.31 to 100 ng/ml with a detection limit of 0.24 ng/ml. The assays showed low reactivities and less than 5% of intraassay and interassay coefficients of variation (CVs), and the average recoveries were between 92 and 97%. Furthermore, the developed method was applied in real samples analysis successfully, and the correlation coefficient with the commercially available OVA kit was 0.9976. Moreover, it was more rapid and sensitive compared with the other methods for testing OVA.
Keywords:Ovalbumin   Chemiluminescence enzyme immunoassay (CLEIA)   Magnetic particles (MPs)
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