Na Guanqiong, Hu Xiaofei, Sun Yaning, Kwee Sharon, Xing Guangxu, Xing Yunrui, Zhang Gaiping
School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, 214122, China.
Key Laboratory for Animal Immunology of the Ministry of Agriculture, Henan Academy of Agricultural Sciences, Zhengzhou, 450002, China.
J Food Sci. 2020 Jun;85(6):1681-1688. doi: 10.1111/1750-3841.15136. Epub 2020 May 17.
Valnemulin (VAL) and tiamulin (TIA) are pleuromutilin antibiotics used primarily for treating bacterial infections in swine or other food animals. Furthermore, VAL and TIA are also employed as feed additives to promote animal growth. However, the illegal use of VAL and TIA could cause a series of hazards to consumers. Here, VAL was designed to be conjugated with bovine serum protein to prepare immunogen. A highly sensitive monoclonal antibody that recognized both VAL and TIA has been successfully produced. Moreover, an immunochromatographic strip assay for rapidly screening VAL and TIA in porcine liver was established with visual detection limits (cutoff values) of 50 and 25 ng/g, respectively. The IC values calculated from the equation of the standard curve were 6.06 and 3.45 ng/g and the limits of detection were 0.96 and 0.29 ng/g for VAL and TIA. According to the recovery experiment results, the test strip exhibited acceptable accuracy and precision. Generally, the proposed strip provided a practical tool for the detection of VAL and TIA. PRACTICAL APPLICATION: We produced a highly sensitive monoclonal antibody and developed an immunoassay strip for simultaneously monitoring TIA and VAL. Additionally it was preliminarily confirmed that the rapid detection tool was suitable for screening TIA and VAL in porcine liver.
伐地米星（VAL）和泰妙菌素（TIA）是截短侧耳素类抗生素，主要用于治疗猪或其他食用动物的细菌感染。此外，VAL和TIA还用作饲料添加剂以促进动物生长。然而，非法使用VAL和TIA会对消费者造成一系列危害。在此，将VAL与牛血清蛋白偶联以制备免疫原。已成功制备出一种能同时识别VAL和TIA的高灵敏度单克隆抗体。此外，还建立了一种用于快速筛查猪肝中VAL和TIA的免疫层析试纸条检测方法，其视觉检测限（临界值）分别为50和25 ng/g。根据标准曲线方程计算出的VAL和TIA的IC值分别为6.06和3.45 ng/g，检测限分别为0.96和0.29 ng/g。根据回收率实验结果，试纸条显示出可接受的准确度和精密度。总体而言，所提出的试纸条为VAL和TIA的检测提供了一种实用工具。实际应用：我们制备了一种高灵敏度单克隆抗体，并开发了一种免疫分析试纸条用于同时监测TIA和VAL。此外，初步证实该快速检测工具适用于筛查猪肝中的TIA和VAL。
