title: An ultrasensitive chemiluminescence immunoassay of chloramphenicol based on gold nanoparticles and magnetic beads.
abstract: A competitive, direct, chemiluminescent immunoassay based on a magnetic beads (MBs) separation and gold nanoparticles (AuNPs) labelling technique to detect chloramphenicol (CAP) has been developed. Horseradish peroxidase (HRP)-labelled anti-CAP monoclonal antibody conjugated with AuNPs and antigen-immobilized MBs were prepared. After optimization parameters of immunocomplex MBs, the IC50 values of chemiluminescence magnetic nanoparticles immunoassay (CL-MBs-nano-immunoassay) were 0.017 µg L(-1) for extract method I and 0.17 µg L(-1) for extract method II. The immunoassay with two extract methods was applied to detect CAP in milk. Comparison of these two extract methods showed that extract method I was advantageous in better sensitivity, in which the sensitivity was 10 times compared to that of extract method II, while extract method II was superior in simple operation, suitable for high throughout screen. The recoveries were 86.7-98.0% (extract method I) and 80.0-103.0% (extract method II), and the coefficients of variation (CVs) were all <15%. The satisfactory recovery with both extract methods and high correlation with traditional ELISA kit in milk system confirmed that the immunomagnetic assay based on AuNPs exhibited promising potential in rapid field screening for trace CAP analysis.
authors.affiliation: Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China.
authors.forename: Xiaoqi
authors.initials: X
authors.lastname: Tao
authors.forename: Haiyang
authors.initials: H
authors.lastname: Jiang
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Yu
authors.forename: Jinghui
authors.initials: J
authors.lastname: Zhu
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
authors.forename: Zhanhui
authors.initials: Z
authors.lastname: Wang
authors.forename: Lanlan
authors.initials: L
authors.lastname: Niu
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Wu
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Shen
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D055910:Antibodies, Immobilized / Q000737:chemistry; D002701:Chloramphenicol / Q000032:analysis*; D006046:Gold / Q000737:chemistry*; D018189:Immunomagnetic Separation / Q000379:methods; D049449:Luminescence; D008163:Luminescent Measurements / Q000379:methods*; D059346:Magnets / Q000737:chemistry; D008892:Milk / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D012680:Sensitivity and Specificity
chemical_list: D000900:Anti-Bacterial Agents; D055910:Antibodies, Immobilized; D002701:Chloramphenicol; D006046:Gold