School of Food Science and Engineering, Engineering Research Center of Bio-process, MOE, Hefei University of Technology, 193 Tunxi Road, Hefei 230009, China.
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
Food Chem. 2018 Sep 15;260:208-212. doi: 10.1016/j.foodchem.2018.04.014. Epub 2018 Apr 7.
We report an aptamer-mediated colorimetric method for sensitive detection of chloramphenicol (CAP). The aptamer of CAP is immobilized by the hybridization with pre-immobilized capture probe in the microtiter plate. The horseradish peroxidase (HRP) is covalently attached to the aptamer by the biotin-streptavidin system for signal production. CAP will preferably bind with aptamer due to the high binding affinity, which attributes to the release of aptamer and HRP and thus, affects the optical signal intensity. Quantitative determination of CAP is successfully achieved in the wide range from 0.001 to 1000 ng/mL with detection limit of 0.0031 ng/mL, which is more sensitive than traditional immunoassays. This method is further validated by measuring the recovery of CAP spiked in two different food matrices (honey and fish). The aptamer-mediated colorimetric method can be a useful protocol for rapid and sensitive screening of CAP, and may be used as an alternative means for traditional immunoassays.
我们报道了一种适体介导的比色法,用于灵敏检测氯霉素(CAP)。CAP 的适体通过与微孔板中预固定的捕获探针杂交固定。辣根过氧化物酶(HRP)通过生物素-链霉亲和素系统共价连接到适体上以产生信号。CAP 由于高结合亲和力而优先与适体结合,这归因于适体和 HRP 的释放,从而影响光学信号强度。通过该方法,在 0.001 至 1000ng/mL 的宽范围内成功实现了 CAP 的定量测定,检测限为 0.0031ng/mL,比传统免疫测定更灵敏。该方法通过测量 CAP 在两种不同食品基质(蜂蜜和鱼)中的加标回收率进一步得到验证。适体介导的比色法可作为快速灵敏筛选 CAP 的有用方案,可作为传统免疫测定的替代手段。