Naik Laxmana, Sharma Rajan, Mann Bimlesh, Lata Kiran, Rajput Y S, Surendra Nath B
Dairy Chemistry Section, ICAR-National Dairy Research Institute, Southern Regional Station, Bengaluru, Karnataka 560030, India.
Dairy Chemistry Division, ICAR-National Dairy Research Institute, Karnal 132001, India.
Food Chem. 2017 Mar 15;219:85-92. doi: 10.1016/j.foodchem.2016.09.090. Epub 2016 Sep 20.
A rapid, semi-quantitative lateral flow assay (LFA) was developed to screen the oxytetracycline (OTC) antibiotics residues in milk samples. In this study a competitive immuno-assay format was established. Colloidal gold nano-particles (GNP) were prepared and used as labelling material in LFA. Polyclonal antibodies were generated against OTC molecule (anti-OTC), purified and the quality was assessed by enzyme linked immuno sorbet assay. For the first time membrane components required for LFA in milk system was optimized. GNP and anti-OTC stable conjugate preparation method was standardized, and then these components were placed over the conjugate pad. OTC coupled with carrier protein was placed on test line; species specific secondary antibodies were placed on the control line of the membrane matrix. Assay was validated by spiking OTC to antibiotic free milk samples and results could be accomplished within 5min. without need of any equipment. The visual detection limit was 30ppb.
开发了一种快速、半定量的侧向流动分析（LFA）方法，用于筛查牛奶样品中的土霉素（OTC）抗生素残留。在本研究中，建立了一种竞争性免疫分析形式。制备了胶体金纳米颗粒（GNP）并将其用作LFA中的标记材料。制备了针对OTC分子的多克隆抗体（抗OTC），进行了纯化，并通过酶联免疫吸附测定评估了其质量。首次对牛奶系统中LFA所需的膜组件进行了优化。标准化了GNP和抗OTC稳定缀合物的制备方法，然后将这些组件放置在缀合垫上。与载体蛋白偶联的OTC放置在检测线上；物种特异性二抗放置在膜基质的对照线上。通过向无抗生素牛奶样品中添加OTC对该分析方法进行了验证，结果可在5分钟内完成，无需任何设备。视觉检测限为30ppb。
