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. Copyright © 2014 John Wiley & Sons, Ltd.