氟苯尼考胶体金免疫层析定量检测方法的建立

Development of Colloidal Gold Lateral Flow Immunoassay for Quantitative Detection of Florfenicol

建立了定量检测氟苯尼考的胶体金免疫层析方法。对胶体金标记抗体时溶液p H和抗体浓度、金标抗体用量、检测线上抗原浓度以及检测时间进行了优化。采用胶体金试纸条读取仪测定试纸条检测线和质控线的信号强度,以标准品的浓度为横坐标,阳性样本和阴性样本的检测线/质控线的信号比值(B_x/B_0)为纵坐标建立标准曲线。结果表明,胶体金免疫层析试纸定量检测氟苯尼考的线性范围为0.1~1.5 ng/m L,检出限为0.08 ng/m L,检测时间为15 min。本方法具有简便、快速和可定量等特点,适于大批量样品的现场筛查。

A method for quantitative detection of florfenicol by colloidal gold lateral flow immunoassay was developed. The experimental conditions including pH value, concentrations of antibody in the process of conjugation between the colloidal gold and antibody, amount of gold-labeled antibody, concentration of the antigen sprayed on test lines （T line） , and detection time were optimized. With a colloidal gold strip reader, the signal intensity of T lines and control lines （ C line） on lateral flow strips was recorded. The T/C ratio of negative control and positive samples was defined as B0 and Bx,and the standard curve was established by plotting the Bx/ B 0 ratio against the concentration of florfenicol. This assay showed a good linear range from 0 .丄 to 丄.5 ng/mL with the limit of detection of 0 . 08 ng/mL, while the result could be obtained within ＇5 min. The result showed that this quantitative method was convenient and rapid, and could be used in screening a large amount of samples on site.