title: A competitive chemiluminescence enzyme immunoassay method for β-defensin-2 detection in transgenic mice.
abstract: A competitive chemiluminescence enzyme immunoassay (CLEIA) method for porcine β-defensin-2 (pBD-2) detection in transgenic mice was established. Several factors that affect detection, including luminol, p-iodophenol and hydrogen peroxide concentrations, as well as pH, were studied and optimized. The linear range of the proposed method for pBD-2 detection under optimal conditions was 0.05-80 ng/mL with a correlation coefficient of 0.9960. Eleven detections of a 30 ng/mL pBD-2 standard sample were performed. Reproducible results were obtained with a relative standard deviation of 3.94%. The limit of detection of the method for pBD-2 was 3.5 pg/mL (3σ). The proposed method was applied to determine pBD-2 expression levels in the tissues of pBD-2 transgenic mice, and compared with LC-MS/MS and quantitative real-time reverse-transcriptase polymerase chain reaction. This suggests that the CLEIA can be used as a valuable method to detect and quantify pBD-2.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China; Division of Animal Infectious Diseases in the State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Yang
authors.forename: Tao
authors.initials: T
authors.lastname: Zhou
authors.forename: Lei
authors.initials: L
authors.lastname: Yu
authors.forename: Wenwen
authors.initials: W
authors.lastname: Tan
authors.forename: Rui
authors.initials: R
authors.lastname: Zhou
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Hu
mesh_terms: D000818:Animals; D007124:Immunoenzyme Techniques / Q000379:methods*; D049449:Luminescence; D051379:Mice; D008822:Mice, Transgenic; D023083:beta-Defensins / Q000032:analysis*
chemical_list: D023083:beta-Defensins
keywords: chemiluminescence enzyme immunoassay; porcine β-defensins-2; transgenic mice